Multiple immunofluorescence technology.
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Animals↗
Biomedical subjects
Cristina Cocco
Publications and source records attributed to Cristina Cocco.
Median eminence dopaminergic nerve terminals: a novel target in autoimmune polyendocrine syndrome?
CONTEXT: Autoantibodies to adenohypophyseal endocrine cells or to vasopressin neurohypophyseal neurons have long been known. Conversely, autoimmune targeting of further hypothalamic-hypophyseal structures, such as the blood-brain barrier-deprived median eminence, has been little studied. OBJECTIVE AND METHODS: We studied a case of autoimmune polyendocrine syndrome type I with GH secretory deficiency, a distinctly rare event in autoimmune polyendocrine syndrome type I. We used rat and bovine tissue substrates to study autoantibodies against hypothalamic-hypophyseal nerve structures and endocrine cells. RESULTS: In the study case, circulating autoantibodies selectively decorated median eminence dopaminergic nerve terminals, as well as pituitary gonadotropes, but not GHRH nerve terminals or pituitary somatotropes. Such autoantibodies appeared de novo in parallel with the onset of GH secretory deficiency, whereas no median eminence labeling was found in patients suffering of idiopathic GH deficiency (n = 7) or in healthy controls (n = 23). CONCLUSIONS: The pathophysiological significance of our patient's autoantibodies remains to be confirmed. Nonetheless, the heterogeneous neuroendocrine structures of the median eminence are pointed out as potential immune targets, relevant to autoimmune polyendocrinopathy, as well as to a wide range of other conditions.
Adolescent↗
Pro-VGF-derived peptides induce penile erection in male rats: possible involvement of oxytocin.
The effect of five peptides derived from the C-terminal portion of rat pro-VGF (VGF(577-617), VGF(588-617), VGF(599-617), VGF(556-576) and VGF(588-597)) on penile erection was studied after injection into the hypothalamic paraventricular nucleus of male rats. VGF(577-617), VGF(588-617), VGF(599-617) and, to a lower extent, VGF(588-597) (0.1-2 microg) induced penile erection episodes in a dose-dependent manner when injected into the paraventricular nucleus, while VGF(556-576) was ineffective. VGF(588-617)-induced penile erection was reduced by nitro(omega)-L-arginine methylester (L-NAME; 20 microg), by morphine (5 microg) and by muscimol (1 microg), but not by dizocilpine [(+)MK-801; 1 microg], nor by cis-flupenthixol (10 microg) given into the paraventricular nucleus 10 min before the VGF peptide. d(CH2)5Tyr(Me)-Orn8-vasotocin (1 microg) effectively reduced VGF(588-617)-induced penile erection when given into the lateral ventricles but not when injected into the paraventricular nucleus. Immunocytochemistry with antibodies specific for the C-terminal nonapeptide sequence of pro-VGF (VGF(609-617)) revealed numerous neuronal fibres and terminals within the paraventricular nucleus, including its parvocellular components. Here, many immunostained neuronal terminals impinged on parvocellular oxytocinergic neurons. The present results show for the first time that certain pro-VGF C-terminus-derived peptides promote penile erection when injected into the paraventricular nucleus and suggest that, within this nucleus, these or closely related pro-VGF-derived peptides may be released to influence sexual function by activating paraventricular oxytocinergic neurons mediating penile erection.
Animals↗
Embedding media for cryomicrotomy: an applicative reappraisal.
We reassess here the formulation of cryoembedding media in connection with recent developments in commercial cryomicrotomes. Water-based solutions of polyvinyl alcohols were our starting media, and each of 2 different polymers (56-98, MW approximately 195000; and 6-98, MW approximately 47000) showed a critical concentration for optimum sectioning. At higher or lower polymer concentrations, wrinkles and folds became apparent in tissue areas of sections, or in the sectioned embedding medium areas between tissues, respectively. Addition of polyethylene glycol (MW 380-420) further facilitated and improved sectioning, resulting in frozen tissue blocks that cut well in the 2 to 100 microm range and further, using disposable blades throughout. Applying a wide temperature differential between tissue specimen (-11 degrees C to -13 degrees C) and cutting knife (-33 degrees C to -35 degrees C), serial adjacent sections were reproducibly obtained at a 2-microm setting, singly or in short ribbons. Embedding media of low and high viscosity were obtained, depending on the polyvinyl alcohol polymer used, and could be applied sequentially for tissue infiltration followed by embedding with precise sample orientation. When required, media were made semisolid by addition of carboxymethylcellulose.
Animals↗
Equipment testing and tuning: the cold-knife cryomicrotome microm HM-560.
A major innovative feature of the Microm HM-560 cryomicrotome is the independent control of specimen and knife temperatures. We used such equipment with a variety of tissues, and fixation and freezing procedures. High-quality sectioning was reproducibly obtained using 1) a low temperature setting for the sectioning blade ("cold knife," about -33 degrees C); 2) a comparatively high temperature for the specimen; and 3) a suitable mounting medium, which would remain solid up to about 0 degrees C. Specimen temperature was set between -8 degrees and -15 degrees C for 4-microm sectioning, higher temperatures (-1 degrees to -8 degrees C) being appropriate when cutting at 10 to 20 microm. Under such conditions, disposable blades were effective throughout, while a modified antiroll plate profile further enhanced usability. After intensive use for almost 3 years, by more than 15 different users, the cryomicrotome is in excellent working order.
Animals↗