No abstract

Teleost sex change is an important model to understand general principles of sexual differentiation and plasticity in the adult brain. The present study is the first to examine the proliferation zones in the adult brain of males, females and sex-changing individuals of a protandrous teleost species (Sparus aurata), by means of 5-bromo-2-deoxyuridine immunocytochemistry. Postnatal neurogenesis in the marine teleost brain was found in ventricular and subventricular areas of the brain that in most cases coincided with the embryonic proliferation zones. The molecular layer of corpus and valvula cerebelli exhibited the highest mitotic activity in the adult brain. High mitotic activity was observed in the hypothalamic, thalamic and telencephalic ventricular areas, as well as the dorsal and ventral rim of the optic tectum. Most of the labeled cells were elongated, indicating the initiation of migratory activity. There were no qualitative differences in the distribution of proliferation zones between the sex phases studied with the exception of the ventricular region of the dorsal hypothalamic area. Volume fraction analysis of the area occupied by the labeled cells suggested that this region included higher densities of newborn cells in the female animals. The proliferation pattern in the adult gilthead sea bream brain is in agreement with the hypothesis of the continuous generation of new cells in the teleost brain. Moreover, our data propose that cell proliferation differences possibly existing in the ventricular region of the dorsal hypothalamus between sexual phases, might be involved in central mechanisms of sexual plasticity in protandrous hermaphrodite teleosts.

The post-injury responses of retinal ganglion cells elicit a number of glial reactions which have not been completely understood. The bilateral pattern of non-neuronal retinal cell proliferation was examined in association with the differential fates of unilaterally injured adult retinal ganglion cells by means of bromodeoxyuridine (BrdU) immunocytochemistry. Lateralization of the glioproliferative events was studied by analysing both the experimental and the uninjured contralateral as well as matched retinas of sham-operated animals. Control adult rat retina included very few BrdU-positive cells within the nerve fibre and ganglion cell layers; however, experimental retinas of degenerating groups exhibited statistically significantly higher densities of newborn cells in most layers. Clusters of labelled cells were found in the inner plexiform layer related to OX-42 staining, indicating their microglial nature. Indeed, double-labelling experiments, after short-term unilateral optic nerve crushing, identified proliferating retinal glial cells in vivo. Both types of glia, astroglial and microglial cells, exhibited BrdU-positive labelling in injured as well as uninjured experimental rat retinas. Moreover, microglial proliferating cells were also identified in explanted retinal pieces after 2 days in culture. Affected and contralateral retinas responded similarly to the unilateral experimental manipulations applied with respect to BrdU labelling. The acute glial responses observed suggest that bilateral glial proliferation might represent a common response related to degeneration events in both retinas, i.e. ipsi- and contralateral to the experimental injury.

It has been reported that neurons generated in the adult brain show sex-specific differences in several brain regions of lower vertebrates and mammals. The present study questioned whether cell proliferation and survival in the adult zebrafish (Danio rerio) cerebellum, the most mitotically active area of adult teleost brain, is sexually differentiated. Adult zebrafish were treated with the thymidine analogue 5'-bromo-2'-deoxyuridine (BrdU) and allowed to survive for 24 h (short-term) and for 21 days (long-term). BrdU immunohistochemistry allowed visualization of cells incorporating BrdU at the S phase of mitosis. At short-term survival, male zebrafish had a higher number of labelled cells at proliferation sites of the molecular layer of corpus cerebelli (CCe) and the granular layer of the caudal lobe of the cerebellum (LCa) than did females. In long-term survival, BrdU-positive cells were found at their final destination, but only the granular layer of the medial division of the valvula cerebelli showed sex-specific differences in the number of labelled cells. This higher mitotic activity in male cerebellum might be related to sex-specific motor behaviour observed in male zebrafish. To investigate the role of programmed cell death, the terminal deoxynucleotidyl-mediated dUTP nick-end-labelling (TUNEL) method was applied. The vast majority of apoptotic figures occurred in the granular cell layer of valvula and CCe, only in a few cases within the BrdU-retaining cells. Apoptosis was found specifically at the sites of the final destination of proliferating cells, indicating that the close relation of cell birth and death might represent a possible plasticity mechanism in the adult zebrafish cerebellum.