The involvement of clock genes in the temporal regulation of the function and lifespan of the corpus luteum (CL) has not been investigated in detail. Immunohistochemistry and real-time quantitative PCR techniques were used to examine the expression of the canonical clock genes: period1, period2, period3, cryptochrome1, cryptochrome2, clock and bmal1, at protein and mRNA levels respectively. The expression of the clock genes was examined in the human CL, cultured luteinised granulosa cells, cultured luteal fibroblast-like cells and the ovine CL. The main findings were that clock genes are expressed in the human and ovine CL; that this expression is manifest at mRNA and protein level in all discernible cell types within the human and ovine CL, and that the pattern of mRNA expression differs between the early luteal phase compared to the late luteal phase. The circadian expression of the clock genes was established in the ovine CL during the late luteal phase and could not be determined in the human CL, although indications from cultured luteinised granulosa cells and luteal fibroblast-like cells suggest that this may also be the case in humans. With the exception of per2, the circadian pattern of clock gene expression emerged in the late luteal phase CL when the early luteal phase CL did not demonstrate circadian clock gene expression. This emergence later in the lifespan of the CL was akin to that observed in embryonic development, where the clock genes are initially non-rhythmic but then acquire circadian rhythmicity with age. In this case, the clock genes have been proposed to perform a non-classical circadian timing role in the timing of embryonic development. The per2 gene was also found to be special, in its loss rather than gain of rhythmic gene expression across the luteal lifespan and in its protein localisation in the cytoplasm of some granulosa-lutein cells. The exceptional behaviour of per2 is consistent with a growing body of evidence supporting its role as a unique clock gene in many respects, able to maintain circadian protein levels in the absence of circadian gene expression, integrating peripheral clock inputs and outputs and acting as a tumour suppressor gene. The CL was also found to be a potential target of melatonin regulation, based on its possession of melatonin MT1 receptors and the timing of circadian cry1 gene expression in the late luteal phase. The expression of cry1 is known to be directly melatonin-induced in the PT and appeared to be similarly activated, downstream of a melatonin signal, in the CL. This supports the evolving view of a hierarchical organisation of the central and peripheral clocks, which are integrated in order to establish information feedback loops that maintain circadian homeostasis, and which can regulate seasonal physiology.

Present study was planned to compare the phenotypic variation of performances traits among commercial (CC) and heritage varieties of turkeys. Information about heritage turkey varieties continues to be limited. In addition, the emerging turkey genome sequence provides a unique opportunity to understand the DNA sequence variation and its associations with performance traits. The turClock, turPer3, turCry1 and turCry2 genes were screened and evaluated for its association with their performance traits. As expected, CC turkeys were superior to heritage birds in performance for most of the traits evaluated. However, heritage turkeys showed better reproductive performances compared to CC turkeys. A total of 41 SNPs were identified from the genes that screened. The haplogroups in the turClock gene were significantly associated with body weight (BW) at 309 d of age, feed conversion ratio (FCR) for 34 - 68 d and 69 - 159 d, egg production and average egg weight (P " 0.05). The haplogroups developed from turPeriod-3 gene were significantly associated with BW at 231 d of age, average daily gain (ADG) for 160 - 231 d, FCR for 69 - 159 d and 160 - 231 d, egg production traits, semen quality traits and plasma melatonin concentration (P " 0.05). In the turCry1 gene, haplogroups were significantly associated with ADG for 35 - 68 d, FCR for 160 - 231 d and 34 - 231 d, egg production and ejaculate volume (P " 0.05). The haplogroups identified from turCry2 gene were significantly associated with BW at 34, 68 and 231 d of age, ADG for 160 - 231 d, FCR for 34 - 68 d, average egg weight (P " 0.05). These findings reveal that phenotypic variation observed in growth and reproductive parameters among turkeys could be used for selecting birds for future breeding programs. DNA sequence variations at the nucleotide and haplotype levels are associated with some of growth, reproductive parameters and plasma melatonin of turkeys. Thus DNA sequence variations that identified of the circadian genes may have some regulatory role in the molecular mechanism of the circadian clock which may affect the circadian rhythm of the animal. / Ph. D.

A melatonina, produzida pela pineal e secretada de maneira cíclica num período de 24 horas, possui ação na sincronização dos ritmos biológicos. Admitindo-se que a expressão circadiana dos clock genes está envolvida na regulação do metabolismo energético e que este padrão oscilatório está sob influência da melatonina, avaliamos o processo de incorporação de glicose em lipídeos no tecido adiposo de ratos controles e pinealectomizados e verificamos a interferência da pinealectomia na expressão dos genes relógio. Os resultados mostram que a pinealectomia alterou a expressão circadiana dos genes Clock, Per2 e Cry1, aumentou a expressão de Rev-erb (Zt 8) e alterou a expressão de Ppar. Ainda, provocou aumento nas concentrações séricas de corticosterona e diminuição nas de leptina. No entanto, o processo de incorporação de glicose em lipídeos não foi alterado. Sendo assim, concluímos que apesar da pinealectomia ter afetado eventos moleculares e metabólicos, quatro semanas de cirurgia ainda é curto para evidenciar repercussões metabólicas mais significativas no animal. / Melatonin, which is produced by pineal gland and secreted in a cyclic fashion over 24 hours, acts in biological rhythms synchronization. Thus, assuming that circadian expression of clock genes is involved in the regulation of energy metabolism and that oscillatory pattern is under influence of melatonin, we analyzed the process of glucose incorporation into lipids in adipose tissue of pinealectomized and controls rats and we determined whether pinealectomy interferes on the expression of clock genes in this tissue. Our results show that pinealectomy alters the circadian pattern of Clock, Per2, Cry1 and Ppar expression, while increases the Rev-erb gene expression (Zt 8). Moreover, the pinealectomy promoted increased corticosterone and decreased leptin plasma levels. However, the incorporation of glucose into lipids over 24 hours was not altered by pinealectomy. Thus, we conclude that despite the pinealectomy have affected molecular and metabolic events, four weeks of surgery is still too short to induce more significant metabolic effects in animals.

A melatonina, produzida pela pineal e secretada de maneira cíclica num período de 24 horas, possui ação na sincronização dos ritmos biológicos. Admitindo-se que a expressão circadiana dos clock genes está envolvida na regulação do metabolismo energético e que este padrão oscilatório está sob influência da melatonina, avaliamos o processo de incorporação de glicose em lipídeos no tecido adiposo de ratos controles e pinealectomizados e verificamos a interferência da pinealectomia na expressão dos genes relógio. Os resultados mostram que a pinealectomia alterou a expressão circadiana dos genes Clock, Per2 e Cry1, aumentou a expressão de Rev-erb (Zt 8) e alterou a expressão de Ppar. Ainda, provocou aumento nas concentrações séricas de corticosterona e diminuição nas de leptina. No entanto, o processo de incorporação de glicose em lipídeos não foi alterado. Sendo assim, concluímos que apesar da pinealectomia ter afetado eventos moleculares e metabólicos, quatro semanas de cirurgia ainda é curto para evidenciar repercussões metabólicas mais significativas no animal. / Melatonin, which is produced by pineal gland and secreted in a cyclic fashion over 24 hours, acts in biological rhythms synchronization. Thus, assuming that circadian expression of clock genes is involved in the regulation of energy metabolism and that oscillatory pattern is under influence of melatonin, we analyzed the process of glucose incorporation into lipids in adipose tissue of pinealectomized and controls rats and we determined whether pinealectomy interferes on the expression of clock genes in this tissue. Our results show that pinealectomy alters the circadian pattern of Clock, Per2, Cry1 and Ppar expression, while increases the Rev-erb gene expression (Zt 8). Moreover, the pinealectomy promoted increased corticosterone and decreased leptin plasma levels. However, the incorporation of glucose into lipids over 24 hours was not altered by pinealectomy. Thus, we conclude that despite the pinealectomy have affected molecular and metabolic events, four weeks of surgery is still too short to induce more significant metabolic effects in animals.

O ritmo diário de atividade é uma característica de todos os organismos vivos, que tem a capacidade de se orientar no tempo e no espaço, e distinguir entre tempo linear e tempo cíclico. O ciclo claro:escuro é um importante indicador circadiano para todos os organismos. O trabalho do relógio circadiano envolve mecanismos de retroalimentação positiva e negativa dos genes CLOCK e BMAL1 (brain and muscle Arnt-like protein 1) que formam um heterodímero, funcionando como fator de transcrição para a expressão dos genes per (period), cry (cryptochrome) e o receptor órfão REV-ERB. Em geral, o ciclo circadiano tem início nas primeiras horas da manhã com a ativação da transcrição de per e cry por CLOCK/BMAL1. A periodicidade do relógio circadiano resulta da combinação entre retroalimentação transcricional positiva e negativa destes genes. Hoje já se sabe que os vertebrados, além do relógio central (NSQ) possuem vários relógios, distribuídos pelo corpo, os chamados relógios periféricos. A resposta ao estímulo luminoso é resultado da interpretação da informação luminosa por diferentes tipos celulares. A molécula fotorreceptora de melanóforos dérmicos embrionários de X. laevis foi denominada melanopsina (Opn4/Opn4). Neste anfíbio, cones e bastonetes, continuam a exibir ritmo circadiano em cultura durante vários dias, e a sua capacidade de se ajustar pelo estímulo luminoso indica a presença do sistema circadiano. Os objetivos deste projeto foram: verificar qual é o padrão de expressão para Opn4, per1, per2, bmal1 e clock em melanóforos de X. laevis submetidos a diferentes fotofases; verificar se a expressão para Opn4, per1, per2 ,bmal1 e clock nos melanóforos de X. laevis é modulada pela melatonina. Opn4, per1, per2 ,bmal1 e clock Dados obtidos no presente estudo demonstram que nesta linhagem celular estes genes apresentam um padrão de expressão aparentemente rítmico, quando estas células são expostas a um ciclo claro:escuro (14C:10E), que difere do padrão obtido quando mantidas em regime de escuro constante. Em geral, estas células mantidas em escuro constante durante 5 dias tendem a apresentar aumento de expressão de RNAm para estes genes e, quando mantidas em escuro constante também durante 5 dias, mas com adição de melatonina por 1h, 24 h antes de sua extração, estes níveis de RNAm tendem a diminuir. Porém, quando comparamos as três situações, podemos observar que a adição da melatonina restaura, em geral, o padrão de expressão dos genes analisados em 14C:10E. O conjunto de resultados, que obtivemos em melanóforos dérmicos de Xenopus laevis, sugere que esta linhagem celular possue características de relógio periférico. / The daily rhythm of activity is a characteristic of all living organisms, which have the ability of to behave accordingly time and space, and distinguish between linear and cyclic time. The dark:light cycle is an important time cue for all organisms. The work of circadian clock involves mechanisms of positive and negative feedback of CLOCK and BMAL1 which as a heterodimer act as a transcription factor for the expression of per (period), cry (cryptochrome) and the orphan receptor REV-ERB. A typical circadian cycle begins in the first hours of daytime, which the activation of the transcription of per and cry by CLOCK/BMAL1. It is well known that the vertebrates, besides the central clock (SCN), have several other clocks distributed by the body, the so called peripheric clock. The responses to light are the result of the interpretation of light signal by several cell types The photoreceptor molecule in the dermal melanophores of X. laevis was denominated melanopsin (Opn4/Opn4). In this amphibian, rods and cones maintain circadian rhythm during several days in culture, and their ability to synchronize by light suggest the presence of a circadian system. The objectives of this project were: verify the expression pattern for Opn4, per1, per2 ,bmal1 e clock in dermal melanophores of X. laevis, under different photo phases; and verify whether the expression for Opn4, per1, per2, bmal1 and clock were modulated by melatonin. Our data show that these genes have a rhythmic pattern expression, when these cells are under a 14L:10D, which is different from the pattern exhibited in constant dark. In general, these cells in constant dark have a higher mRNA expression, and in the same condition, but with melatonin applied for 1h, 24h before the data collect, these mRNA levels are lower. However, when we compared these three different experimental conditions, we observed that melatonin resets, in overall, the expression pattern of 14L:10D. These data, taken together, suggest that Xenous laevis dermal melanophores have characteristics of a peripheric clock.

This thesis focuses on the circadian clock genes and their involvement in the photoperiodic time measurement in the linden bug, Pyrrhocoris apterus. Application of the molecular biology methods enabled us to propose the architecture of circadian clockwork. We also investigated the role of several previously undescribed players in the circadian clock. Furthermore, by using molecular biology methods we focused on the involvement of core circadian clock genes in the photoperiodism.

Background: Shift work has recently been identified as a breast cancer risk factor, where meta-analysis has indicated an approximately 50% increased risk among long-term shift workers. However, additional studies with more comprehensive methods of shift work exposure assessment are needed to capture the diversity of shift patterns. The hypothesized mechanism for this relationship involves chronodisruption (altered circadian rhythms), where increased exposure to light at night during night shifts may decrease production of the cancer-protective hormone melatonin. Further, coordination of circadian rhythms, including melatonin production, is governed by the interactions of a set of central clock genes. Recent studies have suggested that variants in clock genes are associated with cancer risk at multiple sites, including breast cancer, although few studies have considered potential interactions with shift work. Methods: This thesis examined relationships of both shift work and clock gene polymorphisms (and their interactions) with breast cancer risk in a case-control study of 1,142 cases and 1,178 controls. The association between light exposure and melatonin production was also investigated in a longitudinal biomarker study conducted among 123 nurses working a two-day, two-night rotating shift pattern. Results: In the case-control study, an association between breast cancer and ≥30 years of shift work (OR = 2.20, 95%CI = 1.13 – 4.28) was detected, although no relationship with short (0 – 14 years) or medium (15 – 29 years) term shift work was observed. As well, variants in 14 clock-related genes were not associated with breast cancer and there were no apparent interactions with shift work history. In the biomarker study, both peak melatonin levels and daily change in melatonin levels were similar when nurses were working their day and night shifts. Further, on the night shift, a slight inverse relationship between light and change in melatonin was observed (p = 0.04). Conclusions: Taken together, these results contribute to the understanding of both the association between shift work and breast cancer, and the biologic mechanisms underlying this relationship. Since shift work is required for many occupations, understanding the mechanisms through which it impacts breast cancer is important to the development of healthy workplace policy. / Thesis (Ph.D, Community Health & Epidemiology) -- Queen's University, 2012-09-26 20:59:40.209

O ritmo diário de atividade é uma característica de todos os organismos vivos, que tem a capacidade de se orientar no tempo e no espaço, e distinguir entre tempo linear e tempo cíclico. O ciclo claro:escuro é um importante indicador circadiano para todos os organismos. O trabalho do relógio circadiano envolve mecanismos de retroalimentação positiva e negativa dos genes CLOCK e BMAL1 (brain and muscle Arnt-like protein 1) que formam um heterodímero, funcionando como fator de transcrição para a expressão dos genes per (period), cry (cryptochrome) e o receptor órfão REV-ERB. Em geral, o ciclo circadiano tem início nas primeiras horas da manhã com a ativação da transcrição de per e cry por CLOCK/BMAL1. A periodicidade do relógio circadiano resulta da combinação entre retroalimentação transcricional positiva e negativa destes genes. Hoje já se sabe que os vertebrados, além do relógio central (NSQ) possuem vários relógios, distribuídos pelo corpo, os chamados relógios periféricos. A resposta ao estímulo luminoso é resultado da interpretação da informação luminosa por diferentes tipos celulares. A molécula fotorreceptora de melanóforos dérmicos embrionários de X. laevis foi denominada melanopsina (Opn4/Opn4). Neste anfíbio, cones e bastonetes, continuam a exibir ritmo circadiano em cultura durante vários dias, e a sua capacidade de se ajustar pelo estímulo luminoso indica a presença do sistema circadiano. Os objetivos deste projeto foram: verificar qual é o padrão de expressão para Opn4, per1, per2, bmal1 e clock em melanóforos de X. laevis submetidos a diferentes fotofases; verificar se a expressão para Opn4, per1, per2 ,bmal1 e clock nos melanóforos de X. laevis é modulada pela melatonina. Opn4, per1, per2 ,bmal1 e clock Dados obtidos no presente estudo demonstram que nesta linhagem celular estes genes apresentam um padrão de expressão aparentemente rítmico, quando estas células são expostas a um ciclo claro:escuro (14C:10E), que difere do padrão obtido quando mantidas em regime de escuro constante. Em geral, estas células mantidas em escuro constante durante 5 dias tendem a apresentar aumento de expressão de RNAm para estes genes e, quando mantidas em escuro constante também durante 5 dias, mas com adição de melatonina por 1h, 24 h antes de sua extração, estes níveis de RNAm tendem a diminuir. Porém, quando comparamos as três situações, podemos observar que a adição da melatonina restaura, em geral, o padrão de expressão dos genes analisados em 14C:10E. O conjunto de resultados, que obtivemos em melanóforos dérmicos de Xenopus laevis, sugere que esta linhagem celular possue características de relógio periférico. / The daily rhythm of activity is a characteristic of all living organisms, which have the ability of to behave accordingly time and space, and distinguish between linear and cyclic time. The dark:light cycle is an important time cue for all organisms. The work of circadian clock involves mechanisms of positive and negative feedback of CLOCK and BMAL1 which as a heterodimer act as a transcription factor for the expression of per (period), cry (cryptochrome) and the orphan receptor REV-ERB. A typical circadian cycle begins in the first hours of daytime, which the activation of the transcription of per and cry by CLOCK/BMAL1. It is well known that the vertebrates, besides the central clock (SCN), have several other clocks distributed by the body, the so called peripheric clock. The responses to light are the result of the interpretation of light signal by several cell types The photoreceptor molecule in the dermal melanophores of X. laevis was denominated melanopsin (Opn4/Opn4). In this amphibian, rods and cones maintain circadian rhythm during several days in culture, and their ability to synchronize by light suggest the presence of a circadian system. The objectives of this project were: verify the expression pattern for Opn4, per1, per2 ,bmal1 e clock in dermal melanophores of X. laevis, under different photo phases; and verify whether the expression for Opn4, per1, per2, bmal1 and clock were modulated by melatonin. Our data show that these genes have a rhythmic pattern expression, when these cells are under a 14L:10D, which is different from the pattern exhibited in constant dark. In general, these cells in constant dark have a higher mRNA expression, and in the same condition, but with melatonin applied for 1h, 24h before the data collect, these mRNA levels are lower. However, when we compared these three different experimental conditions, we observed that melatonin resets, in overall, the expression pattern of 14L:10D. These data, taken together, suggest that Xenous laevis dermal melanophores have characteristics of a peripheric clock.

No description available.

Submitted by ANGELA CRISTINA DE NICOLA null (angela_bio2006@yahoo.com.br) on 2018-02-07T18:16:28Z No. of bitstreams: 1 Tese Angela.pdf: 2735259 bytes, checksum: bcfc7217ff01dee64fa4ca1839e45c76 (MD5) / Approved for entry into archive by Ana Paula Rimoli de Oliveira null (anapaula@foa.unesp.br) on 2018-02-09T17:42:31Z (GMT) No. of bitstreams: 1 nicola_ml_dr_araca_int.pdf: 2735259 bytes, checksum: bcfc7217ff01dee64fa4ca1839e45c76 (MD5) / Made available in DSpace on 2018-02-09T17:42:31Z (GMT). No. of bitstreams: 1 nicola_ml_dr_araca_int.pdf: 2735259 bytes, checksum: bcfc7217ff01dee64fa4ca1839e45c76 (MD5) Previous issue date: 2017-12-12 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O envelhecimento é considerado processo multidimensional no qual fatores ambientais podem proteger ou, inversamente, agravar seus sinais, de maneira não linear, nos processos fisiológicos e neurocomportamentais. Durante este processo, os ritmos circadianos são interrompidos ou fragmentados com dissociação consequente dos ritmos circadianos do indivíduo e disfunções relacionadas ao relógio circadiano contribuem para o envelhecimento e para patologias a ele relacionadas. O objetivo deste estudo foi averiguar possível alteração temporal do sistema CLOCK no eixo HPG e a relação com às alterações hormonais que caracterizam a periestropausa. Foram utilizadas fêmeas adultas com ciclo estral regular (CD) na fase do diestro e fêmeas senis com ciclo estral irregular e persistência da fase do diestro (IDP). Para análises de expressão gênica dos clock genes Per2, Rev-erbα e Bmal1 no eixo HPG, foram utilizados punchs das regiões do NSQ, onde também foi analisado RNAm de AVP, APO e HMB destes animais, além da adenohipófise e ovários dos quais se extraiu o RNA para confecção do cDNA e realização de qPCR. A determinação da atividade neuronal vasopressinérgica no NSQ foi realizada por imunoistoquíca com dupla marcação para cFos e AVP em tecido previamente fixado com paraformaldeído. A concentração plasmática de gonadotrofinas foi determinada por radioimunoensaio. De modo geral, os animais IDP revelaram alterações no perfil de expressão gênica durante o fotoperíodo, com redução de amplitude, deslocamento/desalinhamento de fase e ausência de antifase. O NSQ de animais IDP apresentou menor expressão de Rev-erbα e maior expressão de RNAm para AVP em relação ao grupo CD. A quantificação relativa de Bmal1 foi semelhante em ambos os grupos e não houve diferenças entre grupos na expressão de Per2. Na APO, animais IDP apresentaram maior expressão de Per2 e menor quantidade de RNAm para Rev-erbα. No HMB observou-se menor expressão para Per2 e Rev-erbα e maior expressão de Bmal1 nas fêmeas IDP. Per2 e Bmal1 na adenohipófise tiveram menor expressão que o gene Rev-erbα no grupo senil e o ovário destes animais revelou maior expressão para Per2 e Rev-erbα, em comparação com os animais CD. As concentrações plasmáticas de FSH foram maiores nas fêmeas com ciclo irregular (2,05 ± 0,44 ng/mL), principalmente durante a fase clara, assim como o LH (0,24 ± 0,07 ng/mL), cujos maiores valores foram encontrados durante a fase escura e com perfil semelhante ao RNAm de AVP. As imunomarcações revelaram alta atividade vasopressinérgica na porção dorsomedial do NSQ das fêmeas IDP. Juntos estes dados permitem concluir que há desarranjo na expressão temporal dos genes Per2, Rev-erbα, Bmal1 que compõem a maquinaria molecular do relógio circadiano, bem como de RNAm para AVP no NSQ, de fêmeas Wistar na periestropausa. Além disso, a maior atividade neuronal vasopressinérgica e a ausência de oscilação de Rev-erbα e Bmal1 no NSQ destes animais, comprometem a correta comunicação do relógio central do NSQ com o eixo HPG, inviabilizando a manutenção da fertilidade feminina e contribuindo para a senescência reprodutiva. / Aging is considered a multidimensional process in which environmental factors can protect or, conversely, aggravate its signals, non-linearly, in physiological and neurobehavioral processes. During this process, circadian rhythms are disrupted or fragmented with consequent dissociation of the individual's circadian rhythms and circadian clock-related dysfunctions contribute to aging and related pathologies. The objective of this study was to investigate possible temporal alteration of the CLOCK system in the HPG axis and the relation with the hormonal changes that characterize periestropause. Adult females with regular estrus cycle in the diestrous phase (RD) and old females with irregular estrous cycle and persistent diestrous phase (IPD). For analyzes of the gene expression of the genes Per2, Rev-erbα and Bmal1 in the HPG axis, punchs from the NSQ regions were used, where AVP, POA and MBH RNAm from these animals were also analyzed, as well as the adenohypophysis and ovaries from which they were extracted the RNA for cDNA production and qPCR performance. The determination of the vasopressinergic neuronal activity in the NSQ was performed by immunohistochemical with double labeling for cFos/AVP in tissue previously fixed with paraformaldehyde. The plasma concentration of gonadotrophins was determined by radioimmunoassay. In general, the IPD animals show alterations in the gene expression profile during the period analyzed, with amplitude reduction, phase shift / misalignment and absence of antiphase. The NSQ of IPD animals presented lower expression of Rev-erbα and higher RNAm expression for AVP than RD group. The relative quantification of Bmal1 was similar in both groups and there were no differences between groups in the expression of Per2. In PAO, IPD animals showed higher expression of Per2 and less amount of RNAm for Rev-erbα. MBH showed lower expression for Per2 and Rev-erbα and higher Bmal1 expression in IPD females. Per2 and Bmal1 in the adenohypophysis had lower expression than the Rev-erbα gene in the old group and the ovary of these animals showed higher expression for Per2 and Rev-erbα, in related to to the RD animals. Plasma concentrations of FSH were higher in females with irregular cycle (2.05 ± 0.44 ng / mL), mainly during the light phase, as well as LH (0.24 ± 0.07 ng / mL) whose values were found during the dark phase and with a profile similar to AVP RNAm. Immunolabeling demonstrated high vasopressinergic activity in the dorsomedial portion of the NSQ of the IPD females. Together these data allow us to conclude that there is a breakdown in the temporal expression of the Per2, Rev-erbα, Bmal1 genes that make up the molecular machinery of the circadian clock, as well as RNAm for AVP in NSQ of Wistar females in peri-masterpause. In addition, the increased vasopressinergic neuronal activity and the absence of Rev-erbα and Bmal1 oscillation in the NSQ of these animals compromise the correct communication of the central clock of the NSQ with the HPG axis, making it impossible to maintain female fertility and contributing to reproductive senescence.