The subject of the thesis is a study of the projections from low threshold oral and face afferents to the cerebral cortex and of descending motor control mechanisms originating in the cerebral cortex or the hypothalamus and influencing the jaw reflexes.Cats anaesthetized with chi oral ose were used for the experiments. Ipsi- and contralateral nerves from the oral cavity and the face were stimulated electrically. Cortical potentials were averaged and recorded. The location of the projections was related to the cytoarchi-tectonic areas of the cerebral cortex. It was found that the afferents projected to separate maximum points in areas 3a, 3b, 5a and 6aß. The projections to areas 3a and 3b were somatotopically organized, but the layout of the projections on the cortex was not facelike.The effect of monopolar anodal stimulation of the cerebral cortex on the monosynaptic jaw closing and the di synaptic jaw opening reflexes was investigated. A sequence of facilitation and inhibition of both reflexes was elicited by cortical stimulation. The effects were of short latency (2.5 ms) and could start with either facilitation or inhibition. The timecourse of the sequence was sinuslike with a period of 10 ms. The largest effect originated in the "sensory" areas 3a and 3b and not in the "motor" areas 4y and 6ag. It is suggested, that a tri gemino-cortico-tri geminai loop via area 3a may function in reflex modulation of jaw movements.The hypothalamic effects on the jaw reflexes were evoked by electrical stimulation in those parts of the hypothalamus, which are w known to generate defence, attack or feeding responses. A tenfold facilitation of the jaw closing reflex and a facilitation followed by almost complete inhibition of the jaw opening reflex were observed in the anaesthetized animal with intact cerebral cortex. The effects remained but were diminished in amplitude after cortical ablation. The descending path was located in the ventral midbrain tegmentum.It is suggested that the observed hypothalamo-tri geminai mechanism may exercise a tonic influence on the trigeminal motoneurones, thereby controlling the set points of the biting force and the rest position. The implications of this hypothesis on the etiology of bruxism and the myofascial pain-dysfunction are discussed. / <p>Härtill 3 rapporter.</p> / digitalisering@umu

Intranuclear rodlets (INRs) are poorly understood intranuclear bodies originally identified within neuronal nuclei on the basis of their unique morphology. Their mechanism of formation, biochemical composition and physiological significance are largely unknown. To gain insight into the molecular regulators of INR formation, mice with a conditional adult β cell-specific knockout of the master regulator of β-cell metabolism, Lkb1 protein kinase (LABKO mice) were studied. The proportion of beta cells containing INRs was significantly reduced in LABKO mice. Further examination ruled out mTOR and Mark2 as downstream effectors of Lkb1 knockout INR phenotype. Instead it identified the mTOR pathway as an independent regulator of INR formation. To investigate INR changes in a pathophysiological context, β cell INRs were examined in two models of human metabolic syndrome: (1) mice maintained on a high-fat diet and (2) leptin-deficient ob/ob mice. Significant INR reduction was observed in both models. Taken together, our results support the view that INR formation in pancreatic β cells is a dynamic and regulated process. The substantial depletion of INRs in LABKO and obese diabetic mice suggests their relationship to β cell function and potential involvement in diabetes pathogenesis. The significance of these findings was further underscored by the demonstration of INRs in human endocrine pancreas, suggesting their potential relevance to the development of metabolic syndrome in humans. The existence of biochemically distinct subtypes of INRs has been suggested by previous reports of differential immunological staining of INRs in neurochemically distinct neuronal populations. Here, a novel variant of INR has been identified that is immunoreactive for the 40kDa huntingtin associated protein and ubiquitin; and evidence was provided for the existence of additional INR subtypes sharing ubiquitin immunoreactivity as a common feature. Selective association of these INRs with melanin concentrating hormone and tyrosine hydroxylase immunoreactive neurons of the hypothalamus and the locus coeruleus was described. It was also demonstrated for the first time that biochemically distinct INR subtypes can co-exist within a single nucleus where they engage in non-random spatial interactions. These findings highlight the biochemical diversity and cell type specific expression of these enigmatic intranuclear structures. On the basis of these findings and previous literature a hypothesis is proposed as to the overall functional significance of INRs in the cell nucleus.

Reproductive senescence in females is a natural part of the aging process. However, the process by which it occurs, and the relative role of each level of the hypothalamic-pituitary-gonadal axis, remains largely unknown. The neural circuitry regulating the hypothalamic axis, including glutamate acting through N-Methyl-D-Aspartate receptors (NMDARs) on GnRH neurons, appears to be key to this process. The NMDAR is tetrameric and composed of an obligatory NR1 subunit together with NR2 subunits. The subunit composition determines the channel kinetics of the receptor and changes through the life span. This dissertation examines the physiological role of the NR2b subunit on LH pulsatile release and LH surge, both important for reproductive function. The expression of NR2b subunits in the anteroventral periventricular (AVPV) nucleus of the hypothalamus was also examined in aging rats. Experiment 1 showed that the NR2b-antagonist, ifenprodil, enhanced pulsatile LH release in estradioltreated females (both age groups). Experiment 2 showed that the LH surge in middle-aged animals was slightly accelerated and that results were dependent upon prior reproductive status of the animals. In Experiment 3, examination of the NR2b-immunoreactive cell population in young, middle-aged, and aged ovariectomized females given vehicle, estradiol, or estradiol with progesterone showed an age-associated decline in NR2b density. However, the immunofluorescent fraction volume of NR1 colocalized with NR2b increased with aging, and that of immunofluorescent fraction volume of NR2b increased with estradiol treatment. This is indicative of the amount of protein expressed in the AVPV. In total, NR2b cell density in the AVPV declines with age, but the amount of NR2b expressed in NR1-positive cells increases, suggesting a larger population of NR2b containing channels. This may translate to age-associated inhibition of GnRH/LH activity, which is relieved with blockade of NR2bcontaining NMDARs. Thus, this dissertation describes a novel way to examine the mechanism by which age-associated changes to neuromodulators of the HPG axis may affect the onset of reproductive senescence. / text

Mice display robust and stereotyped behaviors towards pups: virgin males typically attack pups, while virgin females and sexually experienced males display parental care. I show here that virgin males that are genetically impaired in vomeronasal sensing do not attack pups and are parental, suggesting a key role of the vomeronasal system in controlling male infanticide. In addition, we have identified putative vomeronasal receptors (or receptor groups) for the detection of pup odors, thus uncovering new tools for the molecular and genetic dissection of male infanticide. Further, we have uncovered galanin-expressing neurons in the medial preoptic area (MPOA) as key regulators of male and female parental behavior. Genetic ablation of MPOA galanin- neurons results in dramatic impairment of parental responses in both virgin females and sexually experienced males. In addition, optogenetic activation of these cells in virgin males suppresses infanticide and induces pup grooming. Thus, MPOA galanin-expressing neurons emerge as an essential node of regulation of innate behavior in the hypothalamus that orchestrates male and female parenting while opposing vomeronasal circuits underlying infanticide. Our results provide an entry point for the genetic and circuit-level dissection of mouse parental behavior and its modulation by social experience.

The hypothalamic-pituitary-thyroid axis operates to maintain the circulating concentration of thyroid hormones. Thyrotropin releasing hormone (TRH) is the major hypothalamic messenger controlling the pituitary-thyroid unit. However, the pituitary-thyroid unit responses to various modalities of TRH exposure are not well characterized. Also, interactions between the thyroid axis and other mammalian organ systems, specifically other hormone axes, to maintain the organism's homeostatic state are not well characterized. This work was designed to clarify the response of the pituitary-thyroid unit to TRH and to assess the effects of physiological levels of the rat's primary adrenal cortical hormone, corticosterone, on the thyroid axis. Adult rats were given equal amounts of TRH by intravenous (I.V.) bolus injection or constant intraperitoneal (I.P.) infusion. Both methods resulted in significant increases in plasma thyroid stimulating hormone (TSH), although the time course and peak plasma value varied with the TRH dosage and administration method. Despite the differences in plasma TSH elicited, the thyroid gland responses were similar. Thus, the pituitary is sensitive to the rate and dose of TRH administration. Also, the thyroid is sensitive to plasma levels of TSH but reaches maximum stimulation at submaximal circulating TSH levels. Adrenalectomized female rats, with I.P. and I.V. catheters, were infused with corticosterone (B) to achieve plasma levels within the rat's physiological range. Plasma samples were drawn before and after submaximal TRH (250 ng/100 g Body Weight) administration for assay of TSH and B concentrations. B in the lower half of its physiological range significantly inhibited the increase in plasma TSH observed 10 and 30 minutes after TRH administration. Also, direct stereotaxic infusion of B (50 ng) followed by TRH (1 ng) into the anterior pituitary inhibited the observed increase in plasma TSH. These studies indicate that homeostatic thyroid axis hormone concentrations are maintained by a feedback loop mechanism which is modulated by adrenal hormones. Specifically, physiological levels of corticosterone decrease pituitary sensitivity to TRH in the rat. In addition, the pituitary and thyroid gland exhibit different response patterns to hormonal stimulation.

When an organism is dehydrated, neurons in the Organum vasculosum lamina terminalis (OVL T) sense this variation in plasma osmolality (OSM) and excite magnocellular neurosecretory cells (MNCs) in the supraoptic nucleus (SON) via glutamatergic synapses. The resulting action potential firing of MNCs will result in the secretion of vasopressin (VP) into the blood, which will promote water reabsorption from the kidney. The relationship between plasma VP and OSM (know as the VP-OSM ratio) is known to change in sensitivity during the course of a day. / Lorsqu'un organisme est déshydraté, les neurones dans l'Organum vasculosum lamina terminalis (OVL T) détectent le changement dans l'osmolalité du plasma (OSM) et excitent les cellules magnocellulaires neurosécrétoires (MNCs) dans le noyau supraoptique (SON) avec des synapses glutamatergique. La décharge des potentiels d'action qui survient dans les MNCs génère la sécrétion de vasopressine (VP) dans le sang, qui permettra la réabsorption d'eau au niveau du rein. Le rapport entre la VP et OSM (connu comme étant le rapport VP/OSM) subit des changements de sensibilité durant une journée.

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Diets high in saturated fatty acids are associated with the development of circadian dysregulation, obesity, and type 2 diabetes mellitus. Conversely, unsaturated fatty acids are now known to improve insulin sensitivity, reduce weight gain, and alleviate obesity-induced inflammation. The aforementioned effects of saturated and unsaturated fatty acids have also been identified in the hypothalamus; however, there is a paucity of studies regarding the role of unsaturated fatty acids in circadian rhythms. Therefore, a novel cell model was established to examine the effects of omega-3 fatty acids on circadian rhythms in hypothalamic neurons. The mHypoE-37 cell line expresses Bmal1, Per2, and Rev-erbα in a circadian manner. The saturated fatty acid, palmitate, was found to induce circadian dysregulation of the mHypoE-37 neurons, whereas the unsaturated fatty acid, docosahexaenoic acid, protected against palmitate-induced circadian changes. These studies are the first to identify the potential for unsaturated fatty acids to protect the circadian system.

Diets high in saturated fatty acids are associated with the development of circadian dysregulation, obesity, and type 2 diabetes mellitus. Conversely, unsaturated fatty acids are now known to improve insulin sensitivity, reduce weight gain, and alleviate obesity-induced inflammation. The aforementioned effects of saturated and unsaturated fatty acids have also been identified in the hypothalamus; however, there is a paucity of studies regarding the role of unsaturated fatty acids in circadian rhythms. Therefore, a novel cell model was established to examine the effects of omega-3 fatty acids on circadian rhythms in hypothalamic neurons. The mHypoE-37 cell line expresses Bmal1, Per2, and Rev-erbα in a circadian manner. The saturated fatty acid, palmitate, was found to induce circadian dysregulation of the mHypoE-37 neurons, whereas the unsaturated fatty acid, docosahexaenoic acid, protected against palmitate-induced circadian changes. These studies are the first to identify the potential for unsaturated fatty acids to protect the circadian system.

Intranuclear rodlets (INRs) are poorly understood intranuclear bodies originally identified within neuronal nuclei on the basis of their unique morphology. Their mechanism of formation, biochemical composition and physiological significance are largely unknown. To gain insight into the molecular regulators of INR formation, mice with a conditional adult β cell-specific knockout of the master regulator of β-cell metabolism, Lkb1 protein kinase (LABKO mice) were studied. The proportion of beta cells containing INRs was significantly reduced in LABKO mice. Further examination ruled out mTOR and Mark2 as downstream effectors of Lkb1 knockout INR phenotype. Instead it identified the mTOR pathway as an independent regulator of INR formation. To investigate INR changes in a pathophysiological context, β cell INRs were examined in two models of human metabolic syndrome: (1) mice maintained on a high-fat diet and (2) leptin-deficient ob/ob mice. Significant INR reduction was observed in both models. Taken together, our results support the view that INR formation in pancreatic β cells is a dynamic and regulated process. The substantial depletion of INRs in LABKO and obese diabetic mice suggests their relationship to β cell function and potential involvement in diabetes pathogenesis. The significance of these findings was further underscored by the demonstration of INRs in human endocrine pancreas, suggesting their potential relevance to the development of metabolic syndrome in humans. The existence of biochemically distinct subtypes of INRs has been suggested by previous reports of differential immunological staining of INRs in neurochemically distinct neuronal populations. Here, a novel variant of INR has been identified that is immunoreactive for the 40kDa huntingtin associated protein and ubiquitin; and evidence was provided for the existence of additional INR subtypes sharing ubiquitin immunoreactivity as a common feature. Selective association of these INRs with melanin concentrating hormone and tyrosine hydroxylase immunoreactive neurons of the hypothalamus and the locus coeruleus was described. It was also demonstrated for the first time that biochemically distinct INR subtypes can co-exist within a single nucleus where they engage in non-random spatial interactions. These findings highlight the biochemical diversity and cell type specific expression of these enigmatic intranuclear structures. On the basis of these findings and previous literature a hypothesis is proposed as to the overall functional significance of INRs in the cell nucleus.