Methamphetamine (MA) abuse is a fast growing drug problem, and is the second most widely abused drug world-wide. MA abuse has been linked to the development of symptoms indistinguishable from schizophrenia, referred to as MA psychosis. MA abusing individuals, who most often comprise adolescents and young adults, are 11 times more likely than the general population to develop psychosis. Of further concern is that in utero exposure to MA is also a growing problem, with more women addicts choosing MA as their primary drug. This has significant implications for the neurodevelopment of the child, with subsequent behavioural deficits later in life. Epidemiological studies suggests that in utero or early life MA exposure places a vulnerable individual at greater risk for developing schizophrenia, although this has never been formerly studied either at clinical or pre-clinical level. Animal models of early life adversity, such as post-weaning social isolation rearing (SIR), can assist in understanding the underlying mechanisms in MA abuse and vulnerability to develop MA psychosis. The aim of the current study was to investigate the long term effects of either prenatal (in utero) or early postnatal administration of MA on the development of schizophrenia-like behavioural and neurochemical abnormalities later in life. In the in utero study, pregnant female Wistar rats received either saline (Sal) or MA 5 mg/kg/day for 16 days by subcutaneous (s.c.) injection , starting on prenatal day 13 (PreND-13) up to postnatal day 2 (PostND02). Male offspring were selected for the study. On PostND 21, the animals were weaned and reared under group or isolation reared conditions for 8 weeks. In the early postnatal study, adult male Wistar rats were divided into group reared and SIR conditions from PostND21. Either group received an escalating dose of MA twice a day (0.2 mg/kg – 6 mg/kg s.c.) or Sal for 16 days, from PostND35 to PostND50. Both in utero and early postnatal groups were then subjected to various behavioural tests on PostND78, including assessment of social interaction (SI) and prepulse inhibition (PPI) of acoustic startle. Following behavioural testing, rats were sacrificed and brains snap frozen for later analysis of cortico-striatal monoamine concentrations, superoxide dismutase activity and lipid peroxidation. In the prenatally exposed group no differences in %PPI was observed, although group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including approaching, time together and anogenital sniffing. SIR animals receiving Sal or MA also showed a decrease in rearing. Regarding self-directed behaviours, group reared animals receiving MA and SIR animals receiving Sal or MA showed an increase in self-grooming. Although some disturbances in regional brain monoamines were observed in the frontal cortex and striatum across the groups, this did not reach significance. A significant increase in malondialdehyde was observed in the striatum in group reared animals receiving MA as well as SIR animals receiving Sal or MA, indicating cell damage, possibly of redox origin. In the early postnatal study, %PPI was significantly reduced in group reared animals receiving MA as well as in SIR animals receiving Sal or MA. Group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including rearing, approaching, time together and anogenital sniffing. Regarding self-directed behaviours and locomotor activity, self-grooming and squares crossed was significantly increased in group reared animals receiving MA and SIR animals receiving Sal or MA. A significant increase in DA was evident in the frontal cortex of SIR and grouped housed animals receiving MA. DA in the MA + SIR combination was elevated but not significantly so. None of the treatments affected striatal monoamine levels. In the group reared animals receiving MA as well as the SIR animals receiving Sal or MA, a significant decrease in SOD activity was observed in the frontal cortex, indicating the presence of oxidative stress in this brain region. None of the parameters indicated an additive effect in MA + SIR treated animals. In conclusion, prenatal exposure to MA led to some evidence of late-life behavioural and neurochemical abnormalities akin to schizophrenia, confirming its penchant for psychotogenic effects. However, chronic postnatal MA exposure was more emphatic, being as effective as SIR, a neurodevelopmental model of schizophrenia, in inducing deficits in the above-mentioned behavioural and neurochemical parameters. Thus, early adolescent abuse of MA is a significant risk factor for the later development of schizophrenia or psychosis. However, the risk appeared not to be exacerbated in a population at risk, i.e. in SIR animals. / Thesis (MSc (Pharmacology))--North-West University, Potchefstroom Campus, 2013.

Methamphetamine (MA) abuse is a fast growing drug problem, and is the second most widely abused drug world-wide. MA abuse has been linked to the development of symptoms indistinguishable from schizophrenia, referred to as MA psychosis. MA abusing individuals, who most often comprise adolescents and young adults, are 11 times more likely than the general population to develop psychosis. Of further concern is that in utero exposure to MA is also a growing problem, with more women addicts choosing MA as their primary drug. This has significant implications for the neurodevelopment of the child, with subsequent behavioural deficits later in life. Epidemiological studies suggests that in utero or early life MA exposure places a vulnerable individual at greater risk for developing schizophrenia, although this has never been formerly studied either at clinical or pre-clinical level. Animal models of early life adversity, such as post-weaning social isolation rearing (SIR), can assist in understanding the underlying mechanisms in MA abuse and vulnerability to develop MA psychosis. The aim of the current study was to investigate the long term effects of either prenatal (in utero) or early postnatal administration of MA on the development of schizophrenia-like behavioural and neurochemical abnormalities later in life. In the in utero study, pregnant female Wistar rats received either saline (Sal) or MA 5 mg/kg/day for 16 days by subcutaneous (s.c.) injection , starting on prenatal day 13 (PreND-13) up to postnatal day 2 (PostND02). Male offspring were selected for the study. On PostND 21, the animals were weaned and reared under group or isolation reared conditions for 8 weeks. In the early postnatal study, adult male Wistar rats were divided into group reared and SIR conditions from PostND21. Either group received an escalating dose of MA twice a day (0.2 mg/kg – 6 mg/kg s.c.) or Sal for 16 days, from PostND35 to PostND50. Both in utero and early postnatal groups were then subjected to various behavioural tests on PostND78, including assessment of social interaction (SI) and prepulse inhibition (PPI) of acoustic startle. Following behavioural testing, rats were sacrificed and brains snap frozen for later analysis of cortico-striatal monoamine concentrations, superoxide dismutase activity and lipid peroxidation. In the prenatally exposed group no differences in %PPI was observed, although group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including approaching, time together and anogenital sniffing. SIR animals receiving Sal or MA also showed a decrease in rearing. Regarding self-directed behaviours, group reared animals receiving MA and SIR animals receiving Sal or MA showed an increase in self-grooming. Although some disturbances in regional brain monoamines were observed in the frontal cortex and striatum across the groups, this did not reach significance. A significant increase in malondialdehyde was observed in the striatum in group reared animals receiving MA as well as SIR animals receiving Sal or MA, indicating cell damage, possibly of redox origin. In the early postnatal study, %PPI was significantly reduced in group reared animals receiving MA as well as in SIR animals receiving Sal or MA. Group reared animals receiving MA and SIR animals receiving Sal or MA showed a decrease in social interactive behaviours, including rearing, approaching, time together and anogenital sniffing. Regarding self-directed behaviours and locomotor activity, self-grooming and squares crossed was significantly increased in group reared animals receiving MA and SIR animals receiving Sal or MA. A significant increase in DA was evident in the frontal cortex of SIR and grouped housed animals receiving MA. DA in the MA + SIR combination was elevated but not significantly so. None of the treatments affected striatal monoamine levels. In the group reared animals receiving MA as well as the SIR animals receiving Sal or MA, a significant decrease in SOD activity was observed in the frontal cortex, indicating the presence of oxidative stress in this brain region. None of the parameters indicated an additive effect in MA + SIR treated animals. In conclusion, prenatal exposure to MA led to some evidence of late-life behavioural and neurochemical abnormalities akin to schizophrenia, confirming its penchant for psychotogenic effects. However, chronic postnatal MA exposure was more emphatic, being as effective as SIR, a neurodevelopmental model of schizophrenia, in inducing deficits in the above-mentioned behavioural and neurochemical parameters. Thus, early adolescent abuse of MA is a significant risk factor for the later development of schizophrenia or psychosis. However, the risk appeared not to be exacerbated in a population at risk, i.e. in SIR animals. / Thesis (MSc (Pharmacology))--North-West University, Potchefstroom Campus, 2013.

Methamphetamine (METH) abuse has become a fast growing drug problem that has developed into a global epidemic. In fact, METH is one of the most commonly abused substances with an estimated 35 million abusers worldwide and is said to be the second most popular illicit drug. The Western Province of South Africa has seen a dramatic increase in drug abuse in recent years where METH is the primary or secondary drug of abuse. Interestingly, more than 50% of these individuals are under the age of 20 years. The longer duration of euphoric effects of METH has attracted many users away from cocaine in favour of METH. In addition to the rapid euphoric effect of METH, the direct short-term effects include arousal, reduced fatigue, an increase in blood pressure, reduced appetite as well as sustained attention. Chronic METH abuse may result in debilitating and long-lasting effects that includes mood disorders such as depression. Studies suggest a strong relationship between exposure to adverse environmental factors early in life and the later development of a neuropsychiatric disorder, such as depression. However, these severe consequences do not seem to invoke cessation of the drug. The euphoric and addictive properties of METH causes users to abuse the drug with an increase in frequency and dose, even though it might not have been their original intention. The primary objective of this study was to investigate the effect of early-life administration of METH to stress-sensitive (Flinders Sensitive Line - FSL) and control (Flinders Resistant Line - FRL) rats on depressive-like behaviour and regional brain monoamine levels later in life. The study implemented a sixteen-day period for administration of METH or a vehicle control from postnatal day 19 (PnD19) to postnatal day 34 (PnD34). The latter developmental stage corresponds to pre-adolescence in the rat when neurological development are similar to that seen in human adolescents, and represents the stage when drug abuse is most common in humans. Chronic dosing of METH and saline was performed twice daily at 09:00 and at 15:00. The animals received a sub-cutaneous (SC) escalating dose regimen of METH during the 16 day period (mimicking binging behaviour in humans), with every dose escalating in increments of 0.2 mg/kg from 0.2 mg/kg to 6.0 mg/kg. The study then investigated whether early-life administration of METH would cause depressive-like behaviours directly after the injection period (immediate drug effects before withdrawal on PnD35) or later in life (after the withdrawal period in early adulthood on PnD60). The behavioural effects were assessed in a battery of tests and thereafter the rats were sacrificed and the frontal cortex removed and snap frozen for later analyses of altered neurochemistry. The study demonstrated that chronic METH treatment during pre-adolescence induces significant behavioural changes related to depression in humans directly after the injection period (PnD35) and later in life (PnD60). The animals displayed antidepressant-like behaviour in the forced swim test (FST) before withdrawal, yet a depressogenic effect was observed 25 days post-withdrawal. This effect also seems to be additive to the congenital depressive-like phenotype of FSL rats, suggesting a role for genetic susceptibility. This observation would be in line with the two-hit hypothesis of depression, suggesting that the manifestation of depression will result when a genetic predisposition is followed by an environmental stressor (i.e. METH) later in life. The data suggests a working hypothesis that individuals that already have a predisposition to depression may be more susceptible to developing depression when abusing METH. The fact that the FSL control rats were more immobile than FRL control rats also confirmed the face validity of the FSL genetic rat model of depression. Locomotor activity assessment indicated that METH treatment decreased locomotor activity in FSL and FRL rats compared to their vehicle controls on PnD35 but not on PnD60. It is important to note that the effects observed in locomotor activity could not have contributed to the immobility observed in the FST, confirming that the immobility in the FST indeed reflects psychomotor and not locomotor effects. The study also demonstrated that METH significantly lowers social interaction behaviour in both FRL and FSL rats, both immediately following drug treatment (PnD35) and after withdrawal (PnD60). It is therefore clear that this effect of METH is long-lasting, putatively related to neurodevelopmental effects. In addition, the rats investigated the familiar object for a greater amount of time in the novel object recognition test (nORT) on PnD35 and PnD60 and may be the result of loss of recognition memory for the familiar object. This data confirms that METH results in cognitive memory deficits probably due to sustained adverse neurodevelopmental effects. Neurochemical analyses of the frontal cortex indicated decreased serotonin (5-HT) and norepinephrine (NE) levels on PnD35. METH is widely recognised for its pro-inflammatory effects, while the reduced 5-HT levels observed may have been the result of an increase in circulating pro-inflammatory cytokines. Neurochemical analyses provided thought-provoking data concerning the role of the permissive hypotheses of depression, indicating that dopamine (DA) is most likely not responsible for the behavioural effects observed, at least under the current study conditions, whereas 5-HT is decidedly more involved than expected. The data also suggest that depletion in NE plays a role in the development of depressive-like behaviours following METH exposure. Based on these findings, we propose that disturbances in 5-HT and NE are a crucial mechanism in how METH abuse may precipitate or worsen depressive-like symptoms in individuals who abuse METH. It should be noted that this study does not discard the role of DA in the development of depression after METH exposure, although under the current study conditions it appears that DA does not play a central role. The current study demonstrated that pre-adolescent exposure to METH can reproduce most of the behavioural changes seen in depressed individuals, and that these behavioural data can be used to identify causal neurochemical factors. Environmental stressors such as METH abuse should be regarded as an additional diagnostic criterion and is relevant to an accumulative risk factor hypothesis. Furthermore, although further study is required, the data suggests that early-life exposure to METH may predispose an individual to mood disorders and behavioural abnormalities later in life. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2015

Methamphetamine (METH) abuse has become a fast growing drug problem that has developed into a global epidemic. In fact, METH is one of the most commonly abused substances with an estimated 35 million abusers worldwide and is said to be the second most popular illicit drug. The Western Province of South Africa has seen a dramatic increase in drug abuse in recent years where METH is the primary or secondary drug of abuse. Interestingly, more than 50% of these individuals are under the age of 20 years. The longer duration of euphoric effects of METH has attracted many users away from cocaine in favour of METH. In addition to the rapid euphoric effect of METH, the direct short-term effects include arousal, reduced fatigue, an increase in blood pressure, reduced appetite as well as sustained attention. Chronic METH abuse may result in debilitating and long-lasting effects that includes mood disorders such as depression. Studies suggest a strong relationship between exposure to adverse environmental factors early in life and the later development of a neuropsychiatric disorder, such as depression. However, these severe consequences do not seem to invoke cessation of the drug. The euphoric and addictive properties of METH causes users to abuse the drug with an increase in frequency and dose, even though it might not have been their original intention. The primary objective of this study was to investigate the effect of early-life administration of METH to stress-sensitive (Flinders Sensitive Line - FSL) and control (Flinders Resistant Line - FRL) rats on depressive-like behaviour and regional brain monoamine levels later in life. The study implemented a sixteen-day period for administration of METH or a vehicle control from postnatal day 19 (PnD19) to postnatal day 34 (PnD34). The latter developmental stage corresponds to pre-adolescence in the rat when neurological development are similar to that seen in human adolescents, and represents the stage when drug abuse is most common in humans. Chronic dosing of METH and saline was performed twice daily at 09:00 and at 15:00. The animals received a sub-cutaneous (SC) escalating dose regimen of METH during the 16 day period (mimicking binging behaviour in humans), with every dose escalating in increments of 0.2 mg/kg from 0.2 mg/kg to 6.0 mg/kg. The study then investigated whether early-life administration of METH would cause depressive-like behaviours directly after the injection period (immediate drug effects before withdrawal on PnD35) or later in life (after the withdrawal period in early adulthood on PnD60). The behavioural effects were assessed in a battery of tests and thereafter the rats were sacrificed and the frontal cortex removed and snap frozen for later analyses of altered neurochemistry. The study demonstrated that chronic METH treatment during pre-adolescence induces significant behavioural changes related to depression in humans directly after the injection period (PnD35) and later in life (PnD60). The animals displayed antidepressant-like behaviour in the forced swim test (FST) before withdrawal, yet a depressogenic effect was observed 25 days post-withdrawal. This effect also seems to be additive to the congenital depressive-like phenotype of FSL rats, suggesting a role for genetic susceptibility. This observation would be in line with the two-hit hypothesis of depression, suggesting that the manifestation of depression will result when a genetic predisposition is followed by an environmental stressor (i.e. METH) later in life. The data suggests a working hypothesis that individuals that already have a predisposition to depression may be more susceptible to developing depression when abusing METH. The fact that the FSL control rats were more immobile than FRL control rats also confirmed the face validity of the FSL genetic rat model of depression. Locomotor activity assessment indicated that METH treatment decreased locomotor activity in FSL and FRL rats compared to their vehicle controls on PnD35 but not on PnD60. It is important to note that the effects observed in locomotor activity could not have contributed to the immobility observed in the FST, confirming that the immobility in the FST indeed reflects psychomotor and not locomotor effects. The study also demonstrated that METH significantly lowers social interaction behaviour in both FRL and FSL rats, both immediately following drug treatment (PnD35) and after withdrawal (PnD60). It is therefore clear that this effect of METH is long-lasting, putatively related to neurodevelopmental effects. In addition, the rats investigated the familiar object for a greater amount of time in the novel object recognition test (nORT) on PnD35 and PnD60 and may be the result of loss of recognition memory for the familiar object. This data confirms that METH results in cognitive memory deficits probably due to sustained adverse neurodevelopmental effects. Neurochemical analyses of the frontal cortex indicated decreased serotonin (5-HT) and norepinephrine (NE) levels on PnD35. METH is widely recognised for its pro-inflammatory effects, while the reduced 5-HT levels observed may have been the result of an increase in circulating pro-inflammatory cytokines. Neurochemical analyses provided thought-provoking data concerning the role of the permissive hypotheses of depression, indicating that dopamine (DA) is most likely not responsible for the behavioural effects observed, at least under the current study conditions, whereas 5-HT is decidedly more involved than expected. The data also suggest that depletion in NE plays a role in the development of depressive-like behaviours following METH exposure. Based on these findings, we propose that disturbances in 5-HT and NE are a crucial mechanism in how METH abuse may precipitate or worsen depressive-like symptoms in individuals who abuse METH. It should be noted that this study does not discard the role of DA in the development of depression after METH exposure, although under the current study conditions it appears that DA does not play a central role. The current study demonstrated that pre-adolescent exposure to METH can reproduce most of the behavioural changes seen in depressed individuals, and that these behavioural data can be used to identify causal neurochemical factors. Environmental stressors such as METH abuse should be regarded as an additional diagnostic criterion and is relevant to an accumulative risk factor hypothesis. Furthermore, although further study is required, the data suggests that early-life exposure to METH may predispose an individual to mood disorders and behavioural abnormalities later in life. / MSc (Pharmacology), North-West University, Potchefstroom Campus, 2015