Pituitary growth hormones: further evidence for evolutionary conservatism based on immunochemical studies.
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Immunochemical relatedness of preparations of purified somatotropins (growth hormones) of somatotropins in pituitary extracts from various vertebrate species was investigated by applying an antiserum to a purified somatotropin from a submammalian species, the snapping turtle. With the exception of monkey somatotropin, all mammalian, reptilian, and avian preparations tested showed reactions of identity or near identity by immunodiffusion studies in agar gel. Radioimmunoassay employing labeled rat somatotropin as a tracer and for standards, revealed that these same pituitary preparations gave steep inhibition slopes that were parallel or nearly parallel to each other. Purified somatotropins or somatotropins in pituitary extracts of subreptilian species, including an amphibian and existing primitive fishes, showed partial yet substantial relatedness to mammalian (ray) or reptilian (turtle) somatotropins by both immunodiffusion and radioimmunoassay. Our evidence indicates that the immunochemical relatedness of somatotropins from various vertebrate species appears to be even closer than has been suggested previously, and that a high degree of conservation of structure occurs during evolution. (+info)
M6P/IGF2R imprinting evolution in mammals.
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Imprinted gene identification in animals has been limited to eutherian mammals, suggesting a significant role for intrauterine fetal development in the evolution of imprinting. We report herein that M6P/IGF2R is not imprinted in monotremes and does not encode for a receptor that binds IGF2. In contrast, M6P/IGF2R is imprinted in a didelphid marsupial, the opossum, but it strikingly lacks the differentially methylated CpG island in intron 2 postulated to be involved in imprint control. Thus, invasive placentation and gestational fetal growth are not required for imprinted genes to evolve. Unless there was convergent evolution of M6P/ IGF2R imprinting and receptor IGF2 binding in marsupials and eutherians, our results also demonstrate that these two functions evolved in a mammalian clade exclusive of monotremes. (+info)
Divergent evolution in M6P/IGF2R imprinting from the Jurassic to the Quaternary.
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M6P/IGF2R imprinting first appeared approximately 150 million years ago following the divergence of prototherian from therian mammals. Although M6P/IGF2R is clearly imprinted in opossums and rodents, its imprint status in humans remains ambiguous. It is also still unknown if M6P/IGF2R imprinting was an ancestral mammalian epigenotype or if it evolved convergently. We report herein that M6P/IGF2R is imprinted in Artiodactyla, as it is in Rodentia and Marsupialia, but that it is not imprinted in Scandentia, Dermoptera and Primates, including ringtail lemurs and humans. These results are most parsimonious with a single ancestral origin of M6P/IGF2R imprinting followed by a lineage-specific disappearance of M6P/IGF2R imprinting in Euarchonta. The absence of M6P/IGF2R imprinting in extant primates, due to its disappearance from the primate lineage over 75 million years ago, demonstrates that imprinting at this locus does not predispose to human disease. Moreover, the divergent evolution of M6P/IGF2R imprinting predicts that the success of in vitro embryo procedures such as cloning may be species dependent. (+info)
Distortion product otoacoustic emission and auditory brainstem responses in the echidna (Tachyglossus aculeatus).
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The auditory function of four wild-caught echidnas was measured using distortion product otoacoustic emissions (DPOAEs) and auditory brainstem responses (ABRs). Emission audiograms were constructed by finding the stimulus levels required to produce a criterion emission amplitude at a given stimulus frequency. For an emission amplitude of -10 dB SPL, the median "best threshold" was 28 dB SPL, and this minimum threshold occurred between 4 and 8 kHz for all animals. The relative effective range of auditory function was defined by the frequencies at which the audiogram was 30 dB above its best threshold. For the emission audiograms, the median lower-frequency limit was 2.3 kHz, the upper limit was 18.4 kHz, and the effective range was 2.7 octaves. The audiogram as measured by ABR was also found to be strongly "U" shaped with similar low- and high-frequency limits, i.e., from 1.6 to 13.9 kHz, with an effective range of 3.1 octaves. These results suggest that the echidna has a behavioral hearing sensitivity comparable to that of typical therian mammals (e.g., rabbits and gerbils) but with a significantly narrower frequency range. DPOAE responses were also measured in selected animals as a function of the variation of all four stimulus parameters (frequencies and intensities of both stimulus tones). Overall, the measured emission responses establish that the echidna does have a cochlear amplifier, and that it could be the same type as in therian mammals. The amplification mechanism in the echidna, currently unidentified, clearly operates to frequencies above 20 kHz, higher than the hearing function observed in any birds or reptiles but lower than for typical therian mammals. This raises the possibility that at least some aspects of the mammalian cochlear amplifier developed early in evolution, before the divergence of the monotremes (echidna and platypus) from the mainstream therian mammals (marsupials and placentals). In this respect, the presence or absence of outer hair cell electromotility in monotremes would have important consequences for understanding the function and evolution of the vertebrate inner ear. (+info)
Type I interferon genes from the egg-laying mammal, Tachyglossus aculeatus (short-beaked echidna).
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The type I IFN are an important group of multifunctional cytokines that have, for whatever reason, evolved to a high level of complexity in eutherian mammals such as humans and mice. However, until recently, little was known about the type I IFN systems of the other two groups of extant mammals, the marsupials and the egg-laying monotremes. Preliminary partial type I IFN sequences from the short-beaked echidna were previously found to cluster only with the IFN-beta subtype in phylogenetic analyses, but a lack of sequence information made interpretation of these results tenuous. Here, we report cloning of the full-length genes of representatives from the two previously defined groups of echidna type I IFN by genomic walking PCR. Along with analysis of conserved cysteine placement and promoter elements, phylogenetic analysis incorporating these sequences strongly suggest that the two groups of echidna type I IFN genes are in fact homologous to IFN-alpha and IFN-beta, confirming that the duplication leading to these two major classes of type I IFN occurred prior to the divergence of eutherians and monotremes some 180 million years ago. Thus, even though there are major differences in gene copy number and heterogeneity, separate IFN-alpha and IFN-beta gene families are a feature of the cytokine networks of all three groups of living mammals. (+info)
Distribution of caspase-14 in epidermis and hair follicles is evolutionarily conserved among mammals.
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Caspase-14, a member of the caspase family of cysteine proteases, is almost exclusively expressed in the epidermis. Studies on human and mouse cells and tissues have implicated caspase-14 in terminal differentiation of epidermal keratinocytes and in the formation of the stratum corneum. Here we investigated evolutionary aspects of the role of caspase-14 by analyzing its distribution in the epidermis and hair follicles of representative species of placental mammals, marsupials, and monotremes. Immunocytochemical staining showed that caspase-14 is consistently expressed in the granular and corneous layer of the epidermis of all mammalian species investigated. Ultrastructural analysis using gold-labeled anticaspase-14 antibodies revealed that caspase-14 is associated preferentially with keratin bundles and amorphous material of keratohyalin granules, but is also present in nuclei of transitional cells of the granular layer and in corneocytes. In hair follicles, caspase-14 was diffusely present in cornifying cells of the outer root sheath, in the companion layer, and, most abundantly, in the inner root sheath of all mammalian species here analyzed. In Henle and Huxley layers of the inner root sheath, labeling was seen in nuclei and, more diffusely, among trichohyalin granules of cornifying cells. In summary, the tissue expression pattern and the intracellular localization of caspase-14 are highly conserved among diverse mammalian species, suggesting that this enzyme is involved in a molecular process that appeared early in the evolution of mammalian skin. The association of caspase-14 with keratohyalin and trichohyalin granules may indicate a specific role of caspase-14 in the maturation of these keratinocyte-specific structures. (+info)
Respiration by buried echidnas Tachyglossus aculeatus.
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Short-beaked echidnas have an impressive ability to submerge completely into soil or sand and remain there, cryptic, for long periods. This poses questions about how they manage their respiration, cut off from a free flow of gases. We measured the gradient in oxygen partial pressure (PO2) away from the snouts of buried echidnas and oxygen consumption (VO2) in five individuals under similar conditions, in two substrates with different air-filled porosities (fa). A theoretical diffusion model indicated that diffusion alone was insufficient to account for the flux of oxygen required to meet measured rates of VO2. However, it was noticed that echidnas often showed periodic movements of the anterior part of the body, as if such movements were a deliberate effort to flush the tidal air space surrounding their nostrils. These ;flushing movements' were subsequently found to temporarily increase the levels of interstitial oxygen in the soil around the head region. Flushing movements were more frequent while VO2 was higher during the burrowing process, and also in substrate with lower fa. We conclude that oxygen supply to buried echidnas is maintained by diffusion through the soil augmented by periodic flushing movements, which ventilate the tidal airspace that surrounds the nostrils. (+info)
Body temperature-related structural transitions of monotremal and human hemoglobin.
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In this study, temperature-related structural changes were investigated in human, duck-billed platypus (Ornithorhynchus anatinus, body temperature T(b) = 31-33 degrees C), and echidna (Tachyglossus aculeatus, body temperature T(b) = 32-33 degrees C) hemoglobin using circular dichroism spectroscopy and dynamic light scattering. The average hydrodynamic radius (R(h)) and fractional (normalized) change in the ellipticity (F(obs)) at 222 +/- 2 nm of hemoglobin were measured. The temperature was varied stepwise from 25 degrees C to 45 degrees C. The existence of a structural transition of human hemoglobin at the critical temperature T(c) between 36-37 degrees C was previously shown by micropipette aspiration experiments, viscosimetry, and circular dichroism spectroscopy. Based on light-scattering measurements, this study proves the onset of molecular aggregation at T(c). In two different monotremal hemoglobins (echidna and platypus), the critical transition temperatures were found between 32-33 degrees C, which are close to the species' body temperature T(b). The data suggest that the correlation of the structural transition's critical temperature T(c) and the species' body temperature T(b) is not mere coincidence but, instead, is a more widespread structural phenomenon possibly including many other proteins. (+info)