In "Tone" with dogs: exploring canine musicality.

This study investigates the musical perception skills of dogs through playback experiments. Dogs were trained to distinguish between two different target locations based on a sequence of four ascending or descending notes. A total of 16 dogs of different breeds, age, and sex, but all of them with at least basic training, were recruited for the study. Dogs received training from their respective owners in a suitable environment within their familiar home settings. The training sequence consisted of notes [Do-Mi-Sol#-Do (C7-E7-G7#-C8; Hz frequency: 2093, 2639, 3322, 4186)] digitally generated as pure sinusoidal tones. The training protocol comprised 3 sequential training levels, with each level consisting of 4 sessions with a minimum of 10 trials per session. In the test phase, the sequence was transposed to evaluate whether dogs used relative pitch when identifying the sequences. A correct response by the dog was recorded as 1, while an incorrect response, occurring when the dog chose the opposite zone of the bowl, was marked as 0. Statistical analyses were performed using a binomial test. Among 16 dogs, only two consistently performed above the chance level, demonstrating the ability to recognize relative pitch, even with transposed sequences. This study suggests that dogs may have the ability to attend to relative pitch, a critical aspect of human musicality.

Human social buffer in goats and dogs.

The primary goal of this study was to explore the social buffering effect that humans offer to goats and dogs with limited exposure to human socialization, particularly in situations involving interactions with unfamiliar humans. A total of 13 dogs and 14 goats were selected for the study, all of which had limited prior socialization with humans. Each animal was placed in a testing room with unfamiliar humans for 15 min. Three experimenters aimed to establish a comfortable environment, encouraging social interaction by offering food to the animals and assessing the animals' willingness to accept food and their response to being approached and petted. If both conditions were satisfied, the animals were classified as "social". If one or none of the conditions were met, the animals were classified as "not social". Cortisol levels were measured by collecting blood samples before and after the test. Non-parametric tests together with a GzLM showed that the effect of human social buffering in goats was different in comparison to dogs: goats exhibited higher cortisol levels after the test, while dogs did not show a significant change. Further analysis demonstrated that "social" goats had a lower likelihood of experiencing significant changes in cortisol levels than dogs. Thus, once human interactions are accepted, both species could benefit from social buffering. In summary, this study enhances our understanding of how dogs and goats respond to social interactions with humans in the social buffering effect.

Comparative insights of the neuroanatomical distribution of the gonadotropin-inhibitory hormone (GnIH) in fish and amphibians.

This paper intends to apprise the reader regarding the existing knowledge on the neuroanatomical distribution of GnIH-like peptides in in fish and amphibians in both the adult stage and during ontogenesis. The neuroanatomical distribution of GnIH-like neuropeptides appears quite different in the studied species, irrespective of the evolutionary closeness. The topology of the olfactory bulbs can affect the distribution of neurons producing the GnIH-like peptides, with a tendency to show a more extended distribution into the brains with pedunculate olfactory bulbs. Therefore, the variability of the GnIH-like system could also reflect specific adaptations rather than evolutionary patterns. The onset of GnIH expression was detected very early during development suggesting its precocious roles, and the neuroanatomical distribution of GnIH-like elements showed a generally increasing trend. This review highlights some critical technical aspects and the need to increase the number of species to be studied to obtain a complete neuroanatomical picture of the GnIH-like system.

Puppies in the problem-solving paradigm: quick males and social females.

We report an observational, double-blind study that examined puppies' behaviors while engaged in solving an experimental food retrieval task (food retrieval task instrument: FRTI). The experimental setting included passive social distractors (i.e., the dog's owner and a stranger). The focus was on how the social and physical environment shapes puppies' behaviors according to sex. The dependent variables were the number of tasks solved on an apparatus (Performance Index) and the time required to solve the first task (Speed). Sex and Stress were set as explanatory factors, and Social Interest, FRTI interactions, other behavior, and age as covariates. The main findings were that male puppies solved the first task faster than females. On the other hand, females displayed significantly more social interest and did so more rapidly than males. Males showed delayed task resolution. This study demonstrates sex differences in a problem-solving task in dog puppies for the first time, thus highlighting that sexually dimorphic behavioral differences in problem-solving strategies develop early on during ontogenesis.

When are puppies receptive to emotion-induced human chemosignals? The cases of fear and happiness.

We report an observational, double-blind, experimental study that examines the effects of human emotional odors on puppies between 3 and 6 months and adult dogs (one year and upwards). Both groups were exposed to control, human fear, and happiness odors in a between subjects' design. The duration of all behaviors directed to the apparatus, the door, the owner, a stranger, and stress behaviors was recorded. A discriminant analysis showed that the fear odor activates consistent behavior patterns for both puppies and adult dogs. However, no behavioral differences between the control and happiness odor conditions were found in the case of puppies. In contrast, adult dogs reveal distinctive patterns for all three odor conditions. We argue that responses to human fear chemosignals systematically influence the behaviors displayed by puppies and adult dogs, which could be genetically prefigured. In contrast, the effects of happiness odors constitute cues that require learning during early socialization processes, which yield consistent patterns only in adulthood.

Is this love? Sex differences in dog-owner attachment behavior suggest similarities with adult human bonds.

Sex differences in the behavioral responses of Labrador Retriever dogs in the Strange Situation Test were explored. Behaviors expressed by dogs during seven 3-min episodes were analyzed through a Principal Component Analysis (PCA). The scores of factors obtained were analyzed with a Generalized Linear Mixed Model to reveal the effects of the dog's sex and age and the owner's sex. In Episode 1 (dog and owner) and 5 (dog alone), the PCA identified three and two factors, respectively, which overall explained 68.7% and 59.8% of the variance, with no effect of sex. In Episodes 2 (dog, owner, and stranger), 3 and 6 (dog and stranger), and 4 and 7 (dog and owner), the PCA identified four factors, which overall explained 51.0% of the variance. Effects of sex were found on: Factor 1 (distress), with lower scores obtained by females in Episode 2 and higher in Episode 3; Factor 2 (sociability), which was overall higher in females; Factor 3 (separation-distress), with females, but not males, obtaining higher scores when left with the stranger than when with the owner. Therefore, females were overall more social but seemed more affected than males by the owner's absence. Parallels can be traced between our results and sex differences found in adult human romantic attachment, suggesting that the dog-owner bond has characteristics that are not found in the infant-mother relationship.

The scholar's best friend: research trends in dog cognitive and behavioral studies.

In recent decades, cognitive and behavioral knowledge in dogs seems to have developed considerably, as deduced from the published peer-reviewed articles. However, to date, the worldwide trend of scientific research on dog cognition and behavior has never been explored using a bibliometric approach, while the evaluation of scientific research has increasingly become important in recent years. In this review, we compared the publication trend of the articles in the last 34 years on dogs' cognitive and behavioral science with those in the general category "Behavioral Science". We found that, after 2005, there has been a sharp increase in scientific publications on dogs. Therefore, the year 2005 has been used as "starting point" to perform an in-depth bibliometric analysis of the scientific activity in dog cognitive and behavioral studies. The period between 2006 and 2018 is taken as the study period, and a backward analysis was also carried out. The data analysis was performed using "bibliometrix", a new R-tool used for comprehensive science mapping analysis. We analyzed all information related to sources, countries, affiliations, co-occurrence network, thematic maps, collaboration network, and world map. The results scientifically support the common perception that dogs are attracting the interest of scholars much more now than before and more than the general trend in cognitive and behavioral studies. Both, the changes in research themes and new research themes, contributed to the increase in the scientific production on the cognitive and behavioral aspects of dogs. Our investigation may benefit the researchers interested in the field of cognitive and behavioral science in dogs, thus favoring future research work and promoting interdisciplinary collaborations.

Sex differences in the behavioral responses of dogs exposed to human chemosignals of fear and happiness.

This research focuses on sex differences in the behavioral patterns of dogs when they are exposed to human chemosignals (sweat) produced in happy and fear contexts. No age, breed or apparatus-directed behavior differences were found. However, when exposed to fear chemosignals, dogs' behavior towards their owners, and their stress signals lasted longer when compared to being exposed to happiness as well as control chemosignals. In the happy odor condition, females, in contrast to males, displayed a significantly higher interest to the stranger compared to their owner. In the fear condition, dogs spent more time with their owner compared to the stranger. Behaviors directed towards the door, indicative of exit interest, had a longer duration in the fear condition than the other two conditions. Female dogs revealed a significantly longer door-directed behavior in the fear condition compared to the control condition. Overall the data shows that the effect of exposure to human emotional chemosignals is not sex dependent for behaviors related to the apparatus, the owner or the stress behaviors; however, in the happiness condition, females showed a stronger tendency to interact with the stranger.

Distribution of gonadotropin-inhibitory hormone (GnIH)-like immunoreactivity in the brain and pituitary of the frog (Pelophylax esculentus) during development.

Gonadotropin-inhibitory hormone (GnIH) is a newly discovered hypothalamic RFamide peptide that influences reproduction by regulating brain and pituitary neuroendocrine functions in vertebrates. We report here for the first time, the ontogenetic description of GnIH-like immunoreactivity in the brain, olfactory system, and pituitary of the frog, Pelophylax esculentus. GnIH-like immunoreactive (GnIH-ir) elements were first observed in larvae at stage 24 in the olfactory mucosa, ventral telencephalon, and diencephalon. GnIH-ir-positive staining progressively increased in frequency and intensity during larval growth and other ir perikarya appeared in the medial septum, anterior commissure, dorsal hypothalamus, and posterior tuberculum. A decline in GnIH-ir neurons was seen along the olfactory/vomeronasal/terminal nerve complex in the stages following the pre- and prometamorphosis, while other GnIH-ir neurons showed positivity in the ventromedial surface of the olfactory bulbs and into the habenular nuclei, but the latter are no longer observed in the following stages of development. The anterior-posterior axis in several brain areas, along with the median eminence and pars intermedia of the hypophysis had the appearance of GnIH-ir fibers from early stages, with a progressive increase in the number till metamorphosis in all major subdivisions of the brain. After premetamorphosis, GnIH-ir fibers arising from labeled neurons in the suprachiasmatic nucleus could be seen contacting the ventricular lumen. The transient appearance of GnIH-ir elements in the olfactory system may hint at an olfactory placode origin in the extracranial region. The distribution of GnIH in several brain regions throughout development suggests important involvement of GnIH in multiple brain functions during development.

Multimodal signaling in the visuo-acoustic mismatch paradigm: similarities between dogs and children in the communicative approach.

In this review, we have analyzed the studies on the "mismatch paradigm" or "contrasting paradigm", in which the word indicates an intent that is opposite to the gesture in dogs and children. The studies on children highlighted the importance of the type of gestural messages that, when delivered in a non-ostensive manner, assume less value than the verbal indication; whereas, when more emphasis is given to the gestures, it produces opposite results. Word-trained dogs appear to rely more on words, but in the absence of such specific training, dogs rely more on gestures either in transitive or intransitive actions. Moreover, gestural communication appears easier to generalize, since dogs respond equally well to the gestural messages of familiar persons and strangers, whereas their performance lowers when a stranger provides a vocal message. Visual signals trigger faster responses than auditory signals, whereas verbal indications can at most equal the gestural latencies, but never overcome them. Female dogs appeared to be more proficient in the interpretation of gestural commands, while males performed better in the case of verbal commands. Based on a PRISMA analyses from the Web of Science database, three papers on children and four on dogs were retrieved. Our analyses revealed that gestures are more reliable reference points than words for dogs and children. Future studies should focus on choices related to objects of different values for the subjects. Moreover, the choices of dogs should be compared using known and unknown objects, which might help clarify how familiarity with the objects could differently influence their responses.

Celebrating 50+ years of research on the reproductive biology and endocrinology of the green frog: An overview.

This issue is dedicated to the late Professor Giovanni Chieffi, and this article is an overview of the research on Comparative Endocrinology of reproduction using Rana esculenta (alias Pelophylax esculentus) as a model system. Starting from the early 1970s till today, a large quantity of work have been conducted both in the fields of experimental endocrinology and in the definition of the diffuse neuroendocrine system, with a major focus on the increasing role of regulatory peptides. The various aspects investigated concerned the histological descriptions of principal endocrine glands of the hypothalamic-pituitary-gonadal (HPG) axis, the localization and distribution in the HPG of several different substances (i.e. neurosteroids, hypothalamic peptide hormones, pituitary gonadotropins, gonadal sex steroids, and other molecules), the determination of sex hormone concentrations in both serum and tissues, the hormone manipulations, as well as the gene and protein expression of steroidogenic enzymes and their respective receptors. All together these researches, often conducted considering different periods of the annual reproductive cycle of the green frog, allowed to understand the mechanism of cascade control/regulation of the HPG axis of R. esculenta, characterizing the role of different hormones in the two sexes, and testing the hypotheses about the function of single hormones in different target organs. It becomes evident from the review that, in their simplest form, several features of this species are specular as compared to those of other vertebrate species and that reproduction in this frog species is either under endogenous multi-hormonal control or by a wide array of different factors. Our excursus of this research, spanning almost five decades, shows that R. esculenta has been intensively and successfully used as an animal model in reproductive endocrinology as well as several field studies such as those involving environmental concerns that focus on the effects of endocrine disruptors and other environmental contaminants.

AMPA receptor expression in mouse testis and spermatogonial GC-1 cells: A study on its regulation by excitatory amino acids.

Excitatory amino acids (EAAs) are found present in the nervous and reproductive systems of animals. Numerous studies have demonstrated a regulatory role for Glutamate (Glu), d-aspartate ( d-Asp) and N-methyl- d-aspartate (NMDA) in the control of spermatogenesis. EAAs are able to stimulate the Glutamate receptors, including the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR). Here in, we assess expression of the main AMPAR subunits, GluA1 and GluA2/3, in the mouse testis and in spermatogonial GC-1 cells. The results showed that both GluA1 and GluA2/3 were localized in mouse testis prevalently in spermatogonia. The subunit GluA2/3 was more highly expressed compared with GluA1 in both the testis and the GC-1 cells. Subsequently, GC-1 cells were incubated with medium containing l-Glu, d-Glu, d-Asp or NMDA to determine GluA1 and GluA2/3 expressions. At 30 minutes and 2 hours of incubation, EAA-treated GC-1 cells showed significantly higher expression levels of both GluA1 and GluA2/3. Furthermore, p-extracellular signal-regulated kinase (ERK), p-Akt, proliferating cell nuclear antigen (PCNA), and Aurora B expressions were assayed in l-Glu-, d-Glu-, and NMDA-treated GC-1 cells. At 30 minutes and 2 hours of incubation, treated GC-1 cells showed significantly higher expression levels of p-ERK and p-Akt. A consequent increase of PCNA and Aurora B expressions was induced by l-Glu and NMDA, but not by d-Glu. Our study demonstrates a direct effect of the EAAs on spermatogonial activity. In addition, the increased protein expression levels of GluA1 and GluA2/3 in EAA-treated GC-1 cells suggest that EAAs could activate ERK and Akt pathways through the AMPAR. Finally, the increased PCNA and Aurora B levels may imply an enhanced proliferative activity.

Effects of chemical pollutants on reproductive and developmental processes in Italian amphibians.

It is a widely held belief that environmental contaminants contribute to the decline of amphibian populations. By spending most of their early life in water and later stages on the land, amphibians face a constant risk of exposure to pesticides and other chemical pollutants in both aquatic and terrestrial environments. This review presents an overview of the studies carried out in Italian amphibians to highlight hazardous effects of bioaccumulation of chemical pollutants in juveniles and adults in various contaminated environments. Further, the studies in the laboratory setting assessing the effects of chemical pollutants on reproductive and developmental processes are reported. These studies and their relative references have been summarized in a tabular form. Three prominent contaminant groups were identified: herbicides, insecticides, and fungicides; and only a few works reported the effects of other chemical pollutants. Each pollutant group has been delegated to a section. All through the literature survey, it is seen that interest in this topic in Italy is very recent and sparse, where only a few anuran and caudata species and only some chemical pollutants have been studied.

Gonadotropin-inhibitory hormone (GnIH) in the amphibian brain and its relationship with the gonadotropin releasing hormone (GnRH) system: An overview.

It is well known that the hypothalamic neuropeptide gonadotropin-releasing hormone (GnRH) plays an important role as a primary factor regulating gonadotropin secretion in reproductive processes in vertebrates. The discovery of the presence of a gonadotropin-inhibitory hormone (GnIH) in the brains of birds has further contributed to our understanding of the reproduction control by the brain. GnIH plays a key role in inhibition of reproduction and acts on the pituitary gland and GnRH neurons via a novel G protein-coupled receptor (GPR147). GnIH decreases gonadotropin synthesis and release, thus inhibiting gonadal development and maintenance. The GnRH and GnIH neuronal peptidergic systems are well reported in mammals and birds, but limited information is available regarding their presence and localization in the brains of other vertebrate species, such as reptiles, amphibians and fishes. The aim of this review is to compile and update information on the localization of GnRH and GnIH neuronal systems, with a particular focus on amphibians, summarizing the neuroanatomical distribution of GnIH and GnRH and emphasizing the discovery of GnIH based on RFamide peptides and GnIH orthologous peptides found in other vertebrates and their functional significance.

Proliferative events and apoptotic remodelling in retinal development of common toad (Bufo bufo).

Proliferation and apoptosis are fundamental processes in the development of the retina, and a proper balance of the two phenomena is crucial to correct development of the organ. Despite intense investigation in different vertebrates, only a few studies have analyzed the cell death and the cell division quantitatively in the same species during development. Here we studied the time course of apoptosis and proliferation in the retina of common toad, Bufo bufo, and discuss the findings in an evolutionary perspective. We found cells that were dividing first scattered throughout the retina, then, in later stages, proliferation was confined to the ciliary marginal zone. This pattern was confirmed by the expression of the proliferative marker PCNA. Both proliferation and apoptosis occurred in successive waves, and two apoptotic peaks were detected: one at premetamorphosis 1 and the second at prometamorphosis. PARP-1, a known molecular marker of apoptosis, was used to confirm the data obtained by counting pyknotic nuclei. In summary, proliferative and apoptotic waves display an inverse time-relationship through development, with apoptotic peaks coinciding with low proliferation phases. In a comparative perspective, amphibians follow a developmental pattern similar to other vertebrates, although with different timing.

Neuroanatomical relationships between FMRFamide-immunoreactive components of the nervus terminalis and the topology of olfactory bulbs in teleost fish.

The nervus terminalis (NT) is the most anterior of the vertebrate cranial nerves. In teleost fish, the NT runs across all olfactory components and shows high morphological variability within this taxon. We compare the anatomical distribution, average number and size of the FMRFamide-immunoreactive (ir) NT cells of fourteen teleost species with different positions of olfactory bulbs (OBs) with respect to the ventral telencephalic area. Based on the topology of the OBs, three different neuroanatomical organizations of the telencephalon can be defined, viz., fish having sessile (Type I), pseudosessile (short stalked; Type II) or stalked (Type III) OBs. Type III topology of OBs appears to be a feature associated with more basal species, whereas Types I and II occur in derived and in basal species. The displacement of the OBs is positively correlated with the peripheral distribution of the FMRFamide-ir NT cells. The number of cells is negatively correlated with the size of the cells. A dependence analysis related to the type of OB topology revealed a positive relationship with the number of cells and with the size of the cells, with Type I and II topologies of OBs showing significantly fewer cells and larger cells than Type III. A dendrogram based on similarities obtained by taking into account all variables under study, i.e., the number and size of the FMRFamide-ir NT cells and the topology of OBs, does not agree with the phylogenetic relationships amongst species, suggesting that divergent or convergent evolutionary phenomena produced the olfactory components studied.

Neuroanatomical organization of the brain gonadotropin-inhibitory hormone and gonadotropin-releasing hormone systems in the frog Pelophylax esculentus.

Growing evidence suggests that gonadotropin-inhibitory hormone (GnIH) may play a key role in mediating vertebrate reproduction. GnIH inhibits gonadotropin synthesis and release by decreasing the activity of gonadotropin-releasing hormone (GnRH) neurons as well as by directly regulating gonadotropin secretion from the pituitary. Whereas the presence of GnIH has been widely investigated in various classes of vertebrates, there are very few immunohistochemical reports focusing on GnIH in amphibians. The aim of this study was to assess the presence and neuroanatomical distribution of GnIH-like immunoreactivity in the brain of the anuran amphibian Pelophylax (Rana) esculentus (esculenta) and to explore any potential anatomical relationship with mammalian GnRH-immunoreactive (mGnRH-ir) elements. The GnIH-like immunoreactive (GnIH-ir) system constitutes two distinct subpopulations in the telencephalon and diencephalon, with the highest number of immunoreactive cells located in the preoptic and suprachiasmatic areas. GnIH-ir neurons were also observed in the medial septum, the anterior commissure, the dorsal hypothalamus, the periventricular nucleus of the hypothalamus, and the posterior tuberculum. Scattered GnIH-ir fibers were present in all major subdivisions of the brain but only occasionally in the median eminence. mGnRH-ir neurons were distributed in the mediobasal telencephalon, the medial septal area, and the anterior preoptic area. Double-label immunohistochemistry revealed that the GnRH and GnIH systems coexist and have overlapping distributions at the level of the anterior preoptic area. Some GnIH-ir fibers were in close proximity to mGnRH-ir cell bodies. Our results suggest that both the neuroanatomy and the functional regulation of GnRH release are conserved properties of the hypothalamic GnIH-ir system among vertebrate species.

GnIH and GnRH expressions in the central nervous system and pituitary of Indian major carp, Labeo rohita during ontogeny: An immunocytochemical study.

Gonadotropin-releasing hormone (GnRH) is the major hypothalamic neuropeptide stimulating gonadotropin secretion in vertebrates. In 2000, gonadotropin-inhibitory hormone (GnIH) was discovered as a hypothalamic neuropeptide that inhibits gonadotropin secretion in birds. Subsequent studies have shown that GnIH is present in the brain of other vertebrates. We show for the first time GnIH immunoreactivity in the central nervous system and pituitary during development of Indian major carp, Labeo rohita and compare it with the localization of GnRH. GnIH and GnRH immunoreactivities were observed from the olfactory system to spinal cord throughout development. In the brain, both neuropeptides were localized in the telencephalon, diencephalon including the preoptic area and rhombencephalon. The localization of GnIH and GnRH in the pituitary suggests that these neuropeptides are involved in the regulation of pituitary hormones by an autocrine manner during development. In addition, the presence of GnIH and GnRH in several other brain regions including the olfactory system suggests their involvement in the regulation of other physiological functions.

D-Aspartic acid implication in the modulation of frog brain sex steroid levels.

There is evidence that D-aspartate (D-Asp) modulates sex hormone levels in frog testis by regulating the activity of P450 aromatase (P450 aro), the key enzyme which converts Testosterone (T) in 17ß-Estradiol (E2). Here we report, for the first time, that there is a direct correlation among brain levels of D-Asp, P450 aro, E2 and Estradiol Receptor (ERα) in the male frogs during the reproductive as well as the post-reproductive phases of the breeding cycle, with highest levels being observed in the post-reproductive period. D-Asp i.p. administration to frogs ready for reproduction, induced an increase of brain P450 aro protein expression with concomitant enhancement of both E2 levels and ERα expression; at the same time, brain T levels and Androgen receptor expression decreased. In contrast, in the post-reproductive frogs, D-Asp treatment did not modify any of these parameters. Taken together, these results imply that the regulation of P450 aro expression by D-Asp could be an important step in the control of E2 levels in the frog brain.

A novel telomerase activity and microRNA-21 upregulation identified in a family with palmoplantar keratoderma.

Palmoplantar keratoderma is a set of skin diseases with hyperkeratotic thickening of palms and soles which are characteristic of these heterogeneous group of keratinization disorders. Various genetic mutations, autosomal dominant or recessive, have been identified which may triggerpalmoplantar keratoderma, as KRT9 (Keratin 9), KRT1 (Keratin1), AQP5 (Aquaporin), SERPINB 7 (serine protease inhibitor). The identification of causal mutations is extremely important for the correct diagnosis. Here, we report the case of a family affected from Palmoplantar keratoderma caused by autosomal dominant KRT1 mutations (Unna-Thost disease). Telomerase activation and hTERT expression take a part in the process of cell proliferation and inflammation and microRNAs, as microRNA-21, are emerging as drivers in the regulation of telomerase activity. Here, the patients underwent KRT1 analysis genetic sequence, telomerase activity and miR-21 expression. Beside histopathology assay was performed. The patients presented thickening of the skin on soles of the feet and the palms of the hands, KRT1mutations and showed high expression levels of hTERT and hTR, the gene encoding for the telomeric subunits, and miR-21 (fold change > 1.5 and p value = 0.043), explicating the aberrant proliferation of epidermal layer and the inflammatory state characterizing palmoplantar keratoderma.