Expression of monocyte chemoattractant protein-1 and macrophage colony-stimulating factor in normal and inflamed rat testis

Abstract

Macrophages are numerous in the testicular interstitial tissue under normal conditions and increase during inflammation. The mechanisms involved are poorly characterized. Expression of the macrophage-regulating cytokines monocyte chemoattractant protein (MCP)-1 and macrophage colony-stimulating factor (M-CSF) was examined in the adult rat testis before and after an i.p. injection of an inflammatory stimulus, lipopolysaccharide (LPS). In the normal testis, M-CSF was readily observed using Northern blot and Western blot analysis. In contrast, MCP-1 was not detectable by Northern blot in the normal testis, but was detected using RT-PCR amplification and a sensitive ELISA. After LPS treatment, testicular MCP-1 mRNA and protein expression increased dramatically (up to 400-fold). In-situ hybridization for MCP-1 revealed that production was confined to the interstitium of the inflamed testis, in Leydig cells, peritubular cells, perivascular cells and monocyte-like macrophages, but not in tissue-resident macrophages. Unlike MCP-1, M-CSF mRNA and protein expression in the testis increased only marginally, if at all, after LPS treatment. These results suggest that MCP-1 stimulates the increase in intratesticular macrophages that accompanies LPS-induced inflammation in vivo. Together with M-CSF, MCP-1 may also play a role in maintaining the resident macrophage population of the normal testis.

Professional phagocytic granulocytes of the bony fish gilthead seabream display functional adaptation to testicular microenvironment

“…Thus, mammalian testicular macrophages display a novel cytokine secretion profile compared with peritoneal macrophages [24] and retain their cytotoxic and phagocytic capacities, although they have greatly diminished proinflammatory functions and exhibit immunosuppressive activity [25]. Moreover, the recruitment and maintenance of the resident macrophages are clearly under the control of testicular soluble factors [26], such as macrophage migration inhibitory factor [27] and monocyte chemoattractant protein-1 [28,29]. The testicular sbNLCs are also recruited by testicular soluble factors, show a different cytokine production profile [8], and retain some phagocytic and ROI production capabilities, although they are impaired severely by the testicular microenvironment.…”

Cytokine profiles in the testes of rats treated with lipopolysaccharide reveal localized suppression of inflammatory responses

“…Northern blotting was carried out as described previously (20). The following plasmids were used: 1,264-bp rat IL1␤ cDNA in pMosBlue-T (Amersham Biosciences, Buckinghamshire, UK); 526-bp rat IL10 cDNA in pBlueScript (Stratagene, La Jolla, CA); 793-bp rat TNF␣ cDNA in pBlueScript; and 950-bp rat TGF␤1 cDNA in pMosBlue-T. Glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) cDNA was as described previously (20). ␤-actin cDNA was amplified from testicular mRNA using the primers 5Ј-CTCTTCCAGCCTTC-CTTCCT-3Ј and 5Ј-AAAGCCATGCCAAATGTCTC-3Ј, followed by subcloning into PCR II dual promoter plasmid (Promega, Southhampton, UK) using standard methods.…”

“…The aim of the present study was to examine both proinflammatory and regulatory cytokine production profiles in the whole rat testis in vivo by using our well-characterized LPSinduced inflammation model (19,20,48,49). This model mimics the human response to inflammation and infection of the male reproductive tract, leading to suppression of the hypothalamic-pituitary axis, reduction in androgen production by the Leydig cells, and disruption of the developing germ cells that correlates with the severity of the inflammation (49).…”

“…This model mimics the human response to inflammation and infection of the male reproductive tract, leading to suppression of the hypothalamic-pituitary axis, reduction in androgen production by the Leydig cells, and disruption of the developing germ cells that correlates with the severity of the inflammation (49). We previously have shown that this inhibition of testis function is accompanied by a transcriptional upregulation of the inflammatory mediators IL-1␤ (21), monocyte chemoattractant protein-1 (20), and inducible nitric oxide synthase (48). However, the resident macrophages of the testis appear not to participate in the inflammatory response, and the result is a minor, transient infiltration of circulating monocytes with little or no neutrophil or lymphocyte recruitment (49).…”

Long-term biopermanence of ceramides, cholesteryl esters, and ether-linked triglycerides with very-long-chain PUFA in the cadmium-damaged testis

“…The dystrophic microcalcifications in the interstitium observed at day 30 after both Cd treatments was consistent with a number of these cells also having died in this period, some directly from Cd toxicity, some after phagocytosis of part of the Cd-contaminated tissue debris. In contrast to the irreversibly damaged intra-tubular cells, these cells persisted because they are potentially replaceable, either from preexisting testicular macrophages, which are capable of undergoing mitosis [27], or from new cohorts of monocytes able to be recruited from the circulation [28].…”