Minocycline causes widespread cell death and increases microglial labeling in the neonatal mouse brain.

Minocycline, an antibiotic of the tetracycline family, inhibits microglia in many paradigms and is among the most commonly used tools for examining the role of microglia in physiological processes. Microglia may play an active role in triggering developmental neuronal cell death, although findings have been contradictory. To determine whether microglia influence developmental cell death, we treated perinatal mice with minocycline (45 mg/kg) and quantified effects on dying cells and microglial labeling using immunohistochemistry for activated caspase-3 (AC3) and ionized calcium-binding adapter molecule 1 (Iba1), respectively. Contrary to our expectations, minocycline treatment from embryonic day 18 to postnatal day (P)1 caused a > tenfold increase in cell death 8 h after the last injection in all brain regions examined, including the primary sensory cortex, septum, hippocampus and hypothalamus. Iba1 labeling was also increased in most regions. Similar effects, although of smaller magnitude, were seen when treatment was delayed to P3-P5. Minocycline treatment from P3 to P5 also decreased overall cell number in the septum at weaning, suggesting lasting effects of the neonatal exposure. When administered at lower doses (4.5 or 22.5 mg/kg), or at the same dose 1 week later (P10-P12), minocycline no longer increased microglial markers or cell death. Taken together, the most commonly used microglial "inhibitor" increases cell death and Iba1 labeling in the neonatal mouse brain. Minocycline is used clinically in infant and pediatric populations; caution is warrented when using minocycline in developing animals, or extrapolating the effects of this drug across ages. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 753-766, 2017.

Cellular and molecular mechanisms of sexual differentiation in the mammalian nervous system.

Neuroscientists are likely to discover new sex differences in the coming years, spurred by the National Institutes of Health initiative to include both sexes in preclinical studies. This review summarizes the current state of knowledge of the cellular and molecular mechanisms underlying sex differences in the mammalian nervous system, based primarily on work in rodents. Cellular mechanisms examined include neurogenesis, migration, the differentiation of neurochemical and morphological cell phenotype, and cell death. At the molecular level we discuss evolving roles for epigenetics, sex chromosome complement, the immune system, and newly identified cell signaling pathways. We review recent findings on the role of the environment, as well as genome-wide studies with some surprising results, causing us to re-think often-used models of sexual differentiation. We end by pointing to future directions, including an increased awareness of the important contributions of tissues outside of the nervous system to sexual differentiation of the brain.

Sexually dimorphic patterns of neural activity in response to juvenile social subjugation.

After experiencing juvenile social subjugation (JSS), adult female rats display more severe depression- and anxiety-like behaviors than adult males, suggesting that JSS is encoded in a sex-specific manner. To test this hypothesis, prepubertal rats (P28-33) were subjected to 10 aggressive acts in ≤10 min from an aggressive adult male, a 10 min encounter with a non-aggressive adult male, or to 10 min in an empty, clean cage (handled control) and were sacrificed one hour later. We then used unbiased stereology to estimate the total number and proportion of neurons immunoreactive for the immediate early gene product Fos bilaterally in the basolateral amygdala (BLA), the anterior and posterior subdivisions of the bed nucleus of the stria terminalis, and the paraventricular nucleus of the hypothalamus (PVN). Overall, females' Fos responses were less selective than males'. The BLA in males displayed a selective Fos response to the non-aggressive male, whereas no such selectivity occurred in the BLA of females. Additionally, there were more neurons overall in the left BLA than the right and this lateralization was specific to males. The principal subdivision of the BST (BSTpr) in males responded selectively to JSS, whereas the BSTpr in females responded to both the non-aggressive and aggressive males. We also found that the regional volume and neuron number of the BSTpr is greater in males than in females. Finally, the PVN in males was, like the BLA, selective for the non-aggressive male, whereas none of the experiences elicited a selective response in females. The greater selectivity for non-threatening stimuli in males in three stress-responsive brain regions may be a clue as to why males are less susceptible to the anxiogenic effects of JSS.

Social experience induces sex-specific fos expression in the amygdala of the juvenile rat.

To compare the response of the medial amygdala and central amygdala to juvenile social subjugation (JSS), we used unbiased stereology to quantify the immediate early gene product Fos in prepubertal rats after aggressive or benign social encounters or handling. We estimated the overall number of neurons and the proportion of Fos immunoreactive neurons in the posterodorsal (MePD) and posteroventral medial amygdala (MePV) and the central amygdala (CeA). Experience elicited Fos in a sex- and hemisphere-dependent manner in the MePD. The left MePD was selective for JSS in both sexes, but the right MePD showed a specific Fos response to JSS in males only. In the MePV, irrespective of hemisphere or sex, JSS elicited the greatest amount of Fos, benign social experience elicited an intermediate level, and handling the least. None of the experiential conditions elicited significant levels of Fos in the CeA. We found a previously unreported sex difference in the number of CeA neurons (M>F) that was highly significant and a strong trend toward a sex difference (M>F) in the MePD. These data show that the posterior MeA subnuclei are more responsive to JSS than to benign social interaction, that sex interacts with hemispheric laterality to determine the response of the MePD to JSS and that the MePV responds to social experience and JSS. Taken together, these findings support the hypothesis that juvenile rats process JSS in a sex-specific manner.