Biology subjects

Freites, C. L.

Publications and source records attributed to Freites, C. L..

3 recordsLinked to original sources

bioRxiv2022

Peritubular macrophages phagocyte remains of undifferentiated spermatogonia in mouse testis

The cells involved in spermatogenesis are germ-cells, called spermatogonia, classified as: type A-undifferentiated, type A-intermediate and type B. During the spermatogenesis, more than 75% of the germ-cells undergo apoptosis and most of them are phagocyted by Sertoli cells. Peritubular macrophages in adult mouse testis are macrophages that both stimulate the proliferation and differentiation of undifferentiated spermatogonia in the wall of the seminiferous tubule. They have long processes and ramified appearance that squished between the lateral sides of neighbor myoid cells. We show, that a population of peritubular macrophages, grouped in pairs and activated, phagocyted undifferentiated spermatogonia in apoptosis. In adult mouse testis, 3.3x 105 undifferentiated spermatogonia are in the germinal epithelium and 8,250 of them are in apoptosis. We counted in the testis 2,634.2 {+/-}160 peritubular macrophages with phagocytic activity. If each one phagocyted one undifferentiated spermatogonia in apoptosis, it may indicated that peritubular macrophages phagocyted 31.9 % of the total undifferentiated spermatogonia in apoptosis. According to our knowledges, this is the first time that it is shown that undifferentiated spermatogonia in apoptosis are cleaned by peritubular macrophages. Summary SentenceWe report that peritubular macrophages of adult mouse testis, phagocytic remains of apoptotic undifferentiated spermatogonia. These results show that peritubular macrophages, like Sertoli cells, participate in the remotion of germinal cells in the testis.

cell biology↗

bioRxiv2022

Spatio-temporal dynamics of nuclear CREB1: what does it mean?

In the mammalian pineal gland (PG), cyclic AMP responsive element-binding protein 1 (CREB1) participates in the nocturnal melatonin synthesis that rhythmically modulates physiology and behavior. Phosphorylation of CREB1 present in pinealocyte nuclei is one of the key regulatory steps that drives pineal transcription. The spatio-temporal dynamics of CREB1 itself within PG cell types have not yet been documented. In this study we analyzed total CREB1 via Western blot, and the dynamism of CREB1 nuclear distribution in individual rat pinealocytes using fluorescence immunohistochemistry followed by confocal laser-scanning microscopy and quantitative analysis. Total CREB1 levels remained constant in the PG throughout the light:dark cycle. The distribution pattern of nuclear CREB1 did vary, however, among different PG cells. Pinealocytes emerged as having discrete CREB1 domains within their nucleoplasm that were especially distinct. The number, size, and location of CREB1 foci fluctuated among pinealocytes, within the same PG and among Zeitgeber times. A significantly larger dispersion of CREB1-immunoreactive nuclear sites was found at night. This was not accompanied by changes in the overall transcription activity, which was mostly conserved between the light and dark phases, as shown by the expression of a particular phosphorylated form of the RNA polymerase II (RNAPII-pSer5CTD). Suppression of the nocturnal norepinephrine pulse by chronic bilateral superior cervical ganglionectomy increased CREB1 dispersion in pinealocyte nuclei, as compared to sham-derived cells. In addition, differences in CREB1 distribution were found between sham-operated and non-operated rats at early night. Together, these data suggest that in mature pinealocytes nuclear CREB1 is subjected to a dynamic spatio-temporal distribution. Further studies are necessary to elucidate the underlying mechanisms, including the role of chromatin and interchromatin elements, and to understand the impact of CREB1 reorganization in the pineal transcriptome.

neuroscience↗

bioRxiv2021

Aβ promotes amyloidogenic processing of APP through a Go/Gβγ signaling

Alzheimers disease (AD) is characterized by a cognitive impairment associated to amyloid beta (A{beta}) aggregation and deposition in the brain. A{beta} is generated by sequential cleavage of the amyloid precursor protein (APP) by {beta}-site APP cleaving enzyme 1 (BACE1) and {gamma}-secretase complex. The mechanisms that underlie exacerbated production of A{beta}, favoring its deposition in the brain, is largely unknown. In vitro studies have shown that A{beta} aggregates trigger enhanced production of A{beta} by a yet non described mechanism. Here, we show that in different cell types, including human neurons derived from induced pluripotent stem cells (iPSC), oligomers and fibrils of A{beta} enhance the convergence and interaction of APP and BACE1 in endosomal compartments. We demonstrated a key role of A{beta}-APP/Go/G{beta}{gamma} signaling on the amyloidogenic processing of APP. We show that APP mutants with impaired capacity to bind A{beta} or to activate Go protein, are unable to exacerbate APP and BACE1 colocalization in the presence of A{beta}. Moreover, pharmacological inhibition of G{beta}{gamma} subunits signaling with gallein, abrogate A{beta}-dependent interaction of APP and BACE1 in endosomes preventing {beta}-processing of APP. Collectively, these findings uncover a feed-forward mechanism of amyloidogenesis that might contribute to A{beta} pathology in early stages of AD and suggest that gallein might have clinical relevance.

neuroscience↗