Parvalbumin and calbindin-D28k are calcium-binding proteins, which are considered to be markers for certain populations of GABAergic neurons. Their correct development in the basolateral amygdaloid complex is critical for the proper emotional functioning in adult live of human and animals. Therefore, in this paper we describe the pattern of the morphological differentiation and distribution of immunoreactive elements of the parvalbumin and calbindin-D28k in this complex on the basis of immunohistochemically stained material obtained from embryonic (E20) and postnatal (P0-P90) rat brains. Calbindin-D28k appeared early in the development, already in the prenatal life. At this time immunopositive reaction was visible only in cell bodies. However, during development the population of immunopositive neurons was divided into four types: (1) polygonal; (2) piriform-like; (3) bipolar; and (4) pyramidal-like. Two weeks after birth calbindin-D28k immunoreactivity also appeared in neuropil. First, there were visible calbindin-D28k positive fibers and granules that encircled unstained cell bodies and formed basket-like structures. Subsequently, these granules appeared along proximal parts of unstained dendrites forming, so called 'cartridges'. The distribution of calbindin-D28k positive cells during postnatal life was rather homogenous throughout whole basolateral complex. Intensity of calbindin-D28k immunoreactivity reached mature level on the 21st day after birth.The maturation pattern of parvalbumin immunopositive elements followed the same sequence as calbindin-D28k, but it started much later - since the 17th day after birth and reached mature appearance on the 30th day of life. Contrary to calbindin-D28k, parvalbumin was not homogeneously distributed in the basolateral complex. Originally, parvalbumin was restricted to the magnocellular part of basolateral nucleus but it was finally expressed also in the parvicellular part of basolateral nucleus and the dorsolateral part of lateral nucleus. The differences in development of these two calcium-binding proteins indicate that parvalbumin and calbindin-D28k play diverse roles during development and maturation of the basolateral amygdala.
Neuronal changes in the amygdala basolateral complex were studied during development and maturation in fetal and postnatal rat brains using morphometrical methods. Forty brains of animals of various ages were fixed in formalin, frozen and cut into 25 microm thick sections and stained with cresyl violet or haematoxylin and eosin (H&E). In cresyl violet preparations, the complex appeared for the first time on embryonic day (E)17 and was composed of two homogeneous nuclei lateral and basolateral. On about the seventh postnatal day, each of these nuclei was divided into two parts the first one into the dorsolateral and ventromedial and the second one into the anterior and posterior. Morphometric investigations showed a different increase of the neuronal and nuclear size in various parts of the basolateral complex up to postnatal day (P)14; after that time these parameters did not change significantly. The neuronal density and the total number of neurons stabilized at P7 in all parts of this complex, except for the dorsolateral part of the lateral nucleus in which a 30% decrease of the total number of cells was observed. From P14, in all nuclei under study, the total number of neurons did not change significantly.
Understanding gene expression that is responsive to sensory stimulation is central to elucidate molecular mechanisms underlying neuronal plasticity. In this study we demonstrate two new methods of stimulating whiskers that provide major sensory input to rat neocortex. In the first paradigm, animals were placed on the top of a cylinder and their vibrissae were brushed by hand. In the second paradigm, animals were placed for a brief period of time into a new, wired cage resulting in vibrissae stimulation when they explored the new environment. Both approaches induced c-Fos expression in barrel cortex corresponding to the stimulated vibrissae, especially in layer IV. Layers II/III and V/VI also showed c-Fos induction, but there were no detectable changes in layer VIb. The majority of c-Fos-expressing cells are probably not inhibitory neurons, because they do not show parvalbumin staining. Both paradigms, in contrast to the previous methods, are simple to use and do not require anesthesia, restraint of animals, or elaborate experimental setups.
BackgroundHepsin, (also called TMPRSS1) and TMPRSS3 are type II transmembrane serine proteases (TTSPs) that are involved in cancer progression. TTSPs can remodel extracellular matrix (ECM) and, when dysregulated, promote tumor progression and metastasis by inducing defects in basement membrane and ECM molecules. This study investigated whether the gene and protein expression levels of these TTSPs were associated with breast cancer characteristics or survival.MethodsImmunohistochemical staining was used to evaluate hepsin levels in 372 breast cancer samples and TMPRSS3 levels in 373 samples. TMPRSS1 mRNA expression was determined in 125 invasive and 16 benign breast tumor samples, and TMPRSS3 mRNA expression was determined in 167 invasive and 23 benign breast tumor samples. The gene and protein expression levels were analyzed for associations with breast cancer-specific survival and clinicopathological parameters.ResultsLow TMPRSS1 and TMPRSS3 mRNA expression levels were independent prognostic factors for poor breast cancer survival during the 20-year follow-up (TMPRSS1, P = 0.023; HR, 2.065; 95 % CI, 1.106–3.856; TMPRSS3, P = 0.013; HR, 2.106; 95 % CI, 1.167–3.800). Low expression of the two genes at the mRNA and protein levels associated with poorer survival compared to high levels (log rank P-values 0.015–0.042). Low TMPRSS1 mRNA expression was also an independent marker of poor breast cancer prognosis in patients treated with radiotherapy (P = 0.034; HR, 2.344; 95 % CI, 1.065–5.160). Grade III tumors, large tumor size, and metastasis were associated with low mRNA and protein expression levels.ConclusionsThe results suggest that the TTSPs hepsin and TMPRSS3 may have similar biological functions in the molecular pathology of breast cancer. Low mRNA and protein expression levels of the studied TTSPs were prognostic markers of poor survival in breast cancer.Electronic supplementary materialThe online version of this article (doi:10.1186/s12885-015-1440-5) contains supplementary material, which is available to authorized users.
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