Nesfatin-1 influences the excitability of glucosensing neurons in the hypothalamic nuclei and inhibits the food intake

Nesfatin-1 is a recently discovered neuropeptide that has been shown to decrease food intake after lateral, third, or fourth brain ventricle, cisterna magna administration, or PVN injection in ad libitum fed rats. With regards to the understanding of nesfatin-1 brain sites of action, additional microinjection studies will be necessary to define specific nuclei, in addition to the PVN, responsive to nesfatin-1 to get insight into the differential effects on food intake. In the present study, we evaluated nesfatin-1 action to modulate food intake response upon injection into the specific hypothalamic nuclei (PVN, LHA and VMN) in freely fed rats during the dark phase. We extend previous observations by showing that the nesfatin-1 (50 pmol) injected before the onset of the dark period significantly reduced the 1 to 5 h cumulative food intake in rats cannulated into the PVN, LHA, but not in rats cannulated into the VMN. Glucosensing neurons located in the hypothalamus are involved in glucoprivic feeding and homeostatic control of blood glucose. In order to shed light on the mechanisms by which nesfatin-1 exerts its satiety-promoting actions, we examined the effect of nesfatin-1 on the excitability of hypothalamic glucosensing neurons. Nesfatin-1 excited most of the glucose-inhibited (GI) neurons and inhibited most of the glucose-excited (GE) neurons in the PVN. Of 34 GI neurons in the LHA tested, inhibitory effects were seen in 70.6% (24/34) of GI neurons. The main effects were excitatory after intra-VMN administration of nesfatin-1 in GE neurons (27/35, 77.1%). Thus, our data clearly demonstrate that nesfatin-1 may exert at least a part of its physiological actions on the control of food intake as a direct result of its role in modulating the excitability of glucosensing neurons in the PVN, LHA and VMN.     

Time-related changes in the labeling pattern of motor and sensory neurons innervating the gastrocnemius muscle,as revealed by the retrograde transport of the cholera toxin B subunit

Summary Morphological changes in the motor and sensory neurons in the lumbar spinal cord and the dorsal root ganglia were investigated at different survival times following the injection of the B subunit of cholera toxin (CTB) into the medial gastrocnemius muscle. Unconjugated CTB, visualized immunohistochemically, was found to be retrogradely transported through ventral and dorsal roots to motor neurons in the anterior horn, each lamina in the posterior horn, and ganglion cells in the dorsal root ganglia at L₃–L₆. The largest numbers of labeled motor neurons and ganglion cells were observed 72 h after the injection of CTB. Thereafter, labeled ganglion cells were significantly decreased in number, whereas the amount of labeled motor neurons showed a slight reduction. Motor neurons had extensive dendritic trees filled with CTB, reaching lamina VII and even the pia mater of the lateral funiculus. Labeling was also seen in the posterior horn, but the central and medial parts of laminae II and III had the most extensively labeled varicose fibers, the origin of which was the dorsal root ganglion cells. The results indicate that CTB is taken up by nerve terminals and can serve as a sensitive retrogradely transported marker for identifying neurons that innervate a specific muscle.     

Quantitative protein changes in the hypothalamic neurons of pubertal male rats, castrated neonatally.

The effect of neonatal castration of male rats on the sexual differentiation of the hypothalamus at puberty was studied. Male rats were castrated on days 1, 5 and 7 after birth. Their brains were processed for study on days 83-85. The neurons and cell nuclei of the preoptic area, mediobasal and ventromedial nuclei were assessed for changes in cell and nuclear sizes and dry weight (calculated using interferometric methods). Neonatal castration resulted in size as well as dry weight increase in the neurons of the anterior and mediobasal hypothalamus. The dry weight increased by 34% (P less than 0.001) in the medial preoptic area, by 25% (P less than 0.001) in the arcuate neurons and by 22% (P less than 0.001) in the ventromedial nucleus. The cell nuclei exhibited perceptible weight increase too--in the medial preoptic area 68% (P less than 0.001); 55% in the arcuate neurons (P less than 0.001), and 39% in the ventromedial region. The weight and size increases in neonatally castrated males were equal to those of females of the same age. In rats castrated on day 7, the cell sizes and dry weights of the ventromedial nucleus increased but the cell nuclei exhibited only little change. It is assumed that the changes in the dry weight may be the result of increased synthetic processes in these groups of neurons which are connected with the tonic and cyclic release of gonadotropins. These changes also point to the hypothalamic differentiation shifting to the female type in the absence of the inducing effect of androgens.     

DISTRIBUTION OF GAMMA-AMINOBUTYRIC ACID (GABA) IN THE RAT HYPOTHALAMUS: FUNCTIONAL CORRELATES OF GABA WITH ACTIVITIES OF APPETITE CONTROLLING MECHANISMS

—In the hypothalamus, the highest GABA content (approx. 26 nmol/mm³) was constantly observed in the lateral hypothalamic area (LHA). In other parts of the hypothalamus uneven distribution of GABA was also observed, but areas showing high concentration of GABA did not coincide with the locations of various hypothalamic nuclei. In the LHA, which is known to contain a feeding centre, the anterior part (6.4 and 6.0 mm anterior (A 6.4 and A 6.0) respectively to the vertical zero plane of de Groot) showed a remarkably high content of GABA. The GABA content in the LHA at A 6.4 was decreased during the initial phase of insulin hypoglycemia and, in contrast, showed a significant increase following hyperglycemia induced by alloxan administration. In the ventromedial nucleus (VMH) of the hypothalamus, which is known to contain a satiety centre, the GABA content was increased during the initial phase of insulin hypoglycemia. The results suggest that both certain parts of the LHA and VMH contain or receive GABA-inhibitory neurons and that these neurons may play important physiological roles in controlling functional states of the feeding and satiety centres in the hypothalamus.     

Hypothalamic unit responses to impulses from the hippocampus and mesencephalic reticular formation

The effect of stimulation of the dorsal tegmentum mesencephali and dorsal hippocampus on spontaneous single unit activity of the retrochiasmal (RCA), lateral (LHA), medial dorsal (MHAd) and medial ventral (MHAv) hypothalamic regions was investigated in acute experiments on rabbits. During repetitive mesencephalic stimulation many more neurons were activated in all parts of the hypothalamus than during hippocampal stimulation. During mesencephalic stimulation more neurons were excited than inhibited in LHA, MHAd, and MHAv, but in RCA more were inhibited. During hippocampal stimulation spontaneous activity was inhibited in many more cells than during mesencephalic stimulation. Neurons on which convergence of impulses from the hippocampus and mesencephalon was observed numbered 25% in RCA, 39% in LHA, 46% in MHAd, and 29% in MHAv of the total number recorded. By their action (inhibitory or excitatory) on the hypothalamic neurons the mesencephalon and hippocampus were antagonistic in 60% of cases in RCA, 78% in LHA, and 61% in MHAd.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. Translated from Neirofiziologiya, Vol. 6, No. 5, pp. 489–495, September–October, 1974.     

Studies on regeneration of central nervous system and social ability of the earthworm <Emphasis Type="Italic">Eudrilus eugeniae</Emphasis>

Earthworms are segmented invertebrates that belong to the phylum Annelida. The segments can be divided into the anterior, clitellar and posterior parts. If the anterior part of the earthworm, which includes the brain, is amputated, the worm would essentially survive even in the absence of the brain. In these brain amputee-derived worms, the nerve cord serves as the primary control center for neurological function. In this current work, we studied changes in the expression levels of anti-acetylated tubulin and serotonin as the indicators of neuro-regenerative processes. The data reveal that the blastemal tissues express the acetylated tubulin and serotonin from day four and that the worm amputated at the 7th segment takes 30 days to complete the regeneration of brain. The ability of self-assemblage is one of the specific functions of the earthworm’s brain. The brain amputee restored the ability of self-assemblage on the eighth day.     

Effect of Excitotoxin Lesions in the Medial Prefrontal Cortex on Cortical and Subcortical Catecholamine Turnover in the Rat

Catecholamine turnover in brain areas innervated by dopaminergic neurons was examined 2, 6, and 12 days after bilateral, N-methyl-D-aspartate lesions confined to the rat medial prefrontal cortex. The lesion produced a significant regional increase in the concentration of 3,4-dihydroxyphenylethylamine (DA, dopamine) in both the medial prefrontal cortex and the ventral tegmental area. DA concentrations were increased in the nucleus accumbens on day 6 (128% of control), in the ventral tegmental area on day 2 (130% of control), and in the medial prefrontal cortex on days 2 (145% of control) and 6 (127% of control). The only significant changes in the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC) (197% of control), and in the ratio DOPAC/DA (163% of control) were found in the medial prefrontal cortex on day 6 post-lesion. All parameters had returned to control levels by day 12. DA depletion after the administration of alpha-methyl-p-tyrosine (AMPT) was not significantly different between excitotoxin-lesioned and sham animals on day 6 in all brain regions. Noradrenaline (NA) and 3,4-dihydroxyphenylethyleneglycol concentrations and their ratios, and the depletion of noradrenaline after AMPT were also determined, and the lesion resulted in a significant regional increase in NA in both the nucleus accumbens and the ventral tegmental area. An elevation of NA (147% of control) in the nucleus accumbens was found on day 12. Since the excitotoxin lesion destroys corticofugal efferents from medial prefrontal cortex to the nucleus accumbens, the anterior corpus striatum and the ventral tegmental area, our results provide no evidence for a role of these cortical projections in the regulation of subcortical DA metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cell activation in the hypothalamus after exposure to an antigen (based on c-fos gene expression)

Increase of the c-fos mRNA positive cells number was insignificant in 30 min. following activation of the rat hypothalamic structures with the tetanus toxoid (TT). Elevation of the c-fos mRNA positive cells number occurred in the hypothalamic' posterior (PHA), lateral (LHA), anterior (AHA), areas dorsomedial (DMH), and ventromedial (VMH) nuclei within 2 hours of the TT administration. In 6 hours the c-fos mRNA positive cells number decreased in PHA, LHA, DMH. The c-fos mRNA expression was stable in arquate and supraoptic hypothalamic nuclei following either the TT or saline administration.     

The expression of TRH, its receptors and degrading enzyme is differentially modulated in the rat limbic system during training in the Morris water maze

TRH administration induces arousal, improves cognition, and modulates glutamatergic and cholinergic transmission in hippocampal neurons. To study the possible involvement of TRH neurons in learning and memory processes, gene expression of TRH, its receptors, and pyroglutamyl peptidase (PPII), were measured in limbic regions of water-maze trained rats. Hypothalamus and amygdala showed changes related to the task but not specific to spatial learning while in hippocampus, pro-TRH and TRH-R1 mRNA levels were specifically increased in those animals trained to find a hidden platform. Variation of TRH content and mRNA levels of pro-TRH, TRH-R1, TRH-R2 and PPII are observed in conditions known to activate TRH hypophysiotropic neurons. Changes in some of these parameters could indicate the activation of TRHergic neurons and their possible involvement in some memory related process. Male Wistar rats were immersed (10 times) for 1, 3 or 5 days in a Morris water-maze containing, or not (yoked control) a platform and sacrificed 5, 30 and 60 min after last trial. TRH content and TSH serum levels were determined by radioimmunoassay; mRNA levels of pro-TRH, TRH-R1, TRH-R2, and PPII, by RT-PCR. Exclusive changes due to spatial training were observed in posterior hippocampus of rats trained for 5 days sacrificed after 60min: decreased TRH content and increased mRNA levels of pro-TRH and TRH-R1, particularly in CA3 region (measured by in situ hybridization). The hypothalamus-pituitary axis responded in both yoked and trained animals (increasing serum TSH levels and pro-TRH expression, due to swim-stress); in the amygdala of both groups, pro-TRH expression increased while diminished that of both receptors and PPII. Differential expression of these parameters suggests involvement of TRH hippocampal neurons in memory formation processes while changes in amygdala could relate to TRH anxiolytic role. The differential modulation in anterior and posterior portions of the hippocampus is discussed.     

Structural and metabolic changes in the limbic cortex of the rabbit in response to experimentation with the hypothalamus

By means of the light and electron microscopy, histochemical and cytophotometrical methods the anterior subarea of the cerebral limbic cortex has been studied in 30 experimental and 30 control rabbits. The experimental animals have been given 3, 15 and 30 sessions (1 h per day) of electric irritation (0.05 mA, 50 Hz, 1 msec) on the posterior hypothalamic field (PHF). Twelve rabbits from 30 control animals make an intact group and 18--a group with inactive electrodes inserted into the PHF. After 3 and especially after 15 sessions it has been revealed: in neurons--an acute swelling, edematous alterations, hyperchromatosis and shrinkage, changes in lactate dehydrogenase, succinic dehydrogenase, nicotinamide-adenine-dinucleotide-diaphorase and nicotinamide-adenine-dinucleotide-phosphate-diaphorase activities, in neuroglia--hypertrophy and weakly manifested hyperplasia. After 30 sessions synapses degenerated after the dark type are revealed. More intensive structural and metabolic changes are noted in the middle cytoarchitectonical complex. The changes of the enzymatic activity are considered as certain signs of weakening mitochondrial processes, connected with energy production, increasing glycolysis, decreasing level of the energetic provision of the cytoplasmic synthesis, arising under conditions of a disturbed transneuronal influence on the cortex by the hypothalamus.     

Cerebellar projections to the somatic pretectum in the cat

Neurons in the somatic pretectum receive input from the dorsal column nuclei (DCN) and project to a comparable "somatic" portion of the dorsal accessory nucleus of the inferior olive (DAO). This somatic DAO is reciprocally connected with the anterior interpositus nucleus of the cerebellum. One question that arises is whether this circuitry is further controlled by an output specifically from the anterior interpositus nucleus to the somatic pretectum. Wheatgerm agglutinin conjugated to horseradish peroxidase was injected into various parts of the cat pretectum. Injection sites were interpreted as including the somatic pretectum if neurons in the DCN were retrogradely labeled and if anterograde terminal labeling occurred in somatic DAO. The locations of retrogradely labeled neurons within the deep cerebellar nuclei were then compared in cases in which the injection sites included or excluded the somatic pretectum. In all cases in which the injection site included the somatic pretectum, retrogradely labeled neurons were observed in the anterior interpositus nucleus as well as in the lateral cerebellar nuclei. In some of these cases, neurons in the posterior interpositus and medial nuclei were also labeled. In contrast, in cases in which the pretectal injection site was located outside or at the border of the somatic pretectum, retrogradely labeled neurons were observed only in the lateral, posterior interpositus, and medial nuclei. Thus, the somatic pretectum appears to receive input primarily from neurons in the anterior interpositus nucleus, along with some input from neurons in the lateral nucleus. These results provide additional evidence for a pathway through the DCN in which sequentially processed somatic information has access to and is modulated by cerebellar circuitry. The existence of such a pathway supports the conclusion that neurons in the DCN convey somatic information important not only for cutaneous, kinesthestic, and other bodily sensations, but also for the control of movement.     

Effect of Melatonin Treatment on 24-hour Variations in Hypothalamic Serotonin and Dopamine Turnover During the Preclinical Phase of Freund's Adjuvant Arthritis in Rats

The effect of melatonin treatment on time-of-day variations in hypothalamic serotonin (5-HT) and dopamine (DA) turnover was studied in rats treated with Freund's complete adjuvant (FCA). Animals received s.c. injections of 30 æg of melatonin or vehicle 1 h before lights off for 11 days. On day 10 of treatment, FCA or its vehicle was s.c. injected, and 2 days later, the rats were killed at 6 different time intervals throughout a 24-hour cycle. Hypothalamic 5-HT, 5-hydroxyindole-3-acetic acid (5-HIAA), DA and 3,4-dihydroxyphenylacetic acid (DOPAC) levels were measured by HPLC. 5-HT and DA turnover were estimated from the 5-HIAA/5-HT and DOPAC/DA ratios, respectively. In the anterior hypothalamus, time-of-day variation in 5-HT turnover was suppressed by FCA, an effect counteracted by melatonin treatment. Melatonin also prevented FCA effect on medial hypothalamic 5-HT turnover, while in the posterior hypothalamus, similar daily variations of 5-HT turnover were found in all experimental groups. As far as DA turnover, FCA or melatonin administration suppressed its daily variations in the anterior hypothalamus. Time-of-day variations in medial hypothalamic DA turnover were similar in all groups while only rats treated with melatonin and FCA or its vehicle exhibited significant daily changes of DA turnover in the posterior hypothalamus. Results indicate that melatonin treatment affects partly the 24-hour pattern of variation of hypothalamic 5-HT and DA turnover at an early phase of FCA arthritis in rats.     

Cerebellar Hypoplasia in Gunn Rats: Effects of Bilirubin on the Maturation of Glutamate Decarboxylase, Na,K-ATPase, 2'',3''-Cyclic Nucleotide 3''-Phosphohydrolase, Acetylcholine and Aryl Esterase, Succinate and Lactate Dehydrogenase, and Arylsulfatase Activities

Abstract: We have investigated the postnatal development of cerebellar glutamate decarboxylase (GAD), acetylcholine esterase (AChE), 2′,3′-cyclic nucleotide 3′-phosphohydrolase (CNPase), arylsulfatase, succinate dehydrogenase (SDH), Na,K-ATPase, arylesterase, and lactate dehydrogenase (LDH) activities in homozygous (jj) Gunn rats, comparing them with those in heterozygotes. Also examined were GAD activities in the anterior and posterior parts of the vermis cerebelli on day 13. The specific activities of GAD on the basis of wet weight (g) were lower in jj rats after day 10, and remained unchanged from days 10 to 15 in jj rats with severe hypoplasia. On day 13, GAD on wet weight and protein (mg) bases in the anterior part OfJJ rats was lower than that in the posterior parts of both rats. On the contrary, AChE activities based on wet weight after day 10 and on protein after day 5 in jj rats were significantly elevated. While correlations of GAD on a wet weight basis with the cerebellar wet weight were positive after day 8, those of AChE on wet weight and protein bases were inverse after days 10 and 5, respectively. CNPase and SDH activities based on protein in jj rats were higher after day 15 and showed inverse correlations with the cerebellar wet weight after days 15 and 10, respectively. Arylsulfatase activities on wet weight and protein bases in jj rats, which had a peak on day 20, were significantly high after days 10 and 8, respectively. Arylsulfatase activities by wet weight on days 10 to 20 and by protein after day 8 were inversely correlated with cerebellar wet weight in jj rats. These results suggest that the cerebellar inhibitory neurons, the axons of which make synaptic connections to the target cells from days 10 to 15, are selectively affected by bilirubin inji rats with severe hypoplasia. The enhanced arylsulfatase activity in jj rats on days 10 to 20 may be due to the increased number of lysosomes, suggesting that cell damage by bilirubin followed by cell destruction occurs. A high level of AChE activity in jj rats appears to show a relative increase in density of the mossy fibers in the hypoplastic cerebellum.     

Orexin-A immunoreactive neurons in the rat hypothalamus do not contain neuronal nitric oxide synthase (nNOS)

The lateral hypothalamic area (LHA), a key site involved in the central control of feeding and energy homeostasis, contains populations of neurons that produce the orexin peptides or nitric oxide, two chemical factors that increase food intake. In this study, we used immunohistochemistry to investigate the possibility that rat LHA neurons co-express orexin-A and neuronal nitric oxide synthase (nNOS). The orexin-A and nNOS cell populations in the LHA showed extensive overlap without co-localization, and no evidence of direct anatomic contact was found. The finding that LHA neurons do not co-localize orexin-A and nNOS may suggest that the actions of the orexins and nitric oxide on food intake are mediated via independent mechanisms, however, nitric oxide is a diffusible molecule and could potentially affect the activity of orexin neurons via a non-synaptic mechanism.     

Recognition of deficient nutrient intake in the brain of rat withl-lysine deficiency monitored by functional magnetic resonance imaging,electrophysiologically and behaviorally

Summary Eachl-amino acid (AA) in plasma and brain remains unchanged all day long while normal diet is available. But once restriction ofl-lysine (Lys) was introduced, strong anorexia happened. When Lys deficient diet was offered to rats, their growth were decreased depending upon dietary Lys intake, and they ingested Lys solution in choice quantitatively and both appetite and growth normalized. The recognition site for the deficit in rat's brain was identified by brain oxygenation using a functional MRI that higher signals in the ventromedial hypothalamus and lateral hypothalamic area (LHA) appeared, at 30–50 minutes after Lys injection i.p. and then recovered. Degree of Lys hunger, assayed by bar-pressing (50mg pellet of normal diet/30 presses), was suppressed by Lys micro-injection into the LHA, similar to free Lys ingestion but any other AA never did, suggesting the LHA as recognition site for Lys deficit in rats with Lys deficiency due to AA homeostasis.     

Characteristics of the action of psychostimulants on learning and memory in rats

The effects of intraperitoneal injections of psychostimulant drugs: centrophenoxine (50 mg/kg) and peptide analog of tuftsin TP-1 (300 micrograms/kg) on the processes of elaboration and retention of conditioned reflex have been experimentally studied. The animals were trained with food reward in 30 trials per day. Psychostimulant drugs or saline were injected after the 10th trial. The elaboration of conditioned reflex was continued 30 min later. The retention was tested 24 h, 7 and 30 days later. It was established that both drugs induce an increase in memory traces stability during 30 days. These findings provide direct evidence that psychostimulant drugs, centrophenoxine and peptide substance TP-1, when injected during consolidation phase can enhance memory storage processes.     

Effects of restricted feeding on the activity of hypothalamic Orexin (OX)-A containing neurons and OX2 receptor mRNA level in the paraventricular nucleus of rats

We have examined the effects of 3 weeks of food restriction on both the activity of neurons containing hypothalamic orexin (OX)-A and the level of OX receptor type 2 (OX2R) mRNA in the paraventricular nucleus (PVN) of rats. Double immunohistochemistry was used to examine the expression of OX-A and Fos in the lateral hypothalamic area (LHA), and in situ hybridization histochemistry was used to measure levels of OX2R mRNA in the PVN. After the period of restricted feeding, 20-30% of OX-A-containing neurons exhibited Fos-like immunoreactivity (LI). The distribution of OX-A-LI/Fos-LI cells in the food-restricted rats was similar to that observed in glucose-deprived rats after intracerebroventricular (icv) administration of 2-deoxy-D-glucose (2-DG). In addition, 3 weeks of food restriction caused a significant decrease in the expression of the OX2R gene in the parvocellular division of the PVN. These results suggest that the activation of OX-A-containing neurons induced by restricted feeding may be involved in neuroendocrine responses to food restriction.     

Modifications of Neuronal Activity in the Medial Vestibular Nucleus during Long-Term Vibration

The impulse background activity (BA) of neurons of the rat medial vestibular nucleus (MVN) was subjected to computer analysis, and its modifications related to long-term vibration were studied. It was shown that following 5 days of 2-h-long vibration sessions, statistically significant changes in some basic characteristics of BA generated by MVN neurons were observed. More than a twofold increase in the mean BA frequency and substantial shifts practically of all statistical BA parameters were found after 10 days of vibration. Following a 15-day-long vibration, MVN neurons showed a clear-cut tendency to restore control values of the BA indices, which probably was related to adaptation processes.     

Metabolic hormones,grazing behaviour,offspring physical distance and productive response of beef cow grazing at two herbage allowances

Herbage allowance (HA) management during gestation–lactation cycle of cows grazing native grasslands improves pregnancy rates and calves’ weaning weight records. Those improvements were associated with greater herbage mass, and better body condition score (BCS) and metabolic status of the cows, which could affect grazing and maternal behaviour, particularly when temporary suckling restriction (TSR) and flushing (FL) is applied. The objective of this study was to evaluate the effect of HA during the gestation–lactation period on cows’ and calves’ liveweight (LW), BCS, milk yield, insulin, IGF-1 and leptin concentrations, as well as the proportion of diurnal grazing activities (grazing, ruminating, walking and idling), and maternal behaviour from −10 to 70 days relative to TSR (day 0 = initiation of TSR at 86 ± 10 days relative to calving). Thirty-three primiparous Hereford cows were allocated to HA treatments during gestation and lactation, which annually averaged 2.5 (low = LHA) and 4 kg DM/kg LW (high = HHA). The LW and BCS of cows did not differ during −10 to 50 days but were greater in HHA than LHA at the end of the study (P < 0.05). Concentrations of IGF-1 were greater in HHA compared to LHA, while insulin and leptin did not differ. Grazing was lower in HHA than LHA, and the opposite occurred with ruminating and idling (P < 0.05). Cow–calf physical distance was greater in LHA cows compared to HHA (P < 0.05) and increased greatly in the former group after FL, while this increase was lower and later in HHA cows. Milk yield was greater in HHA (P < 0.05), and calves’ weight did not differ between treatments from day −10 to 35, but was greater in HHA compared to LHA from 45 days until the end of the study. Thus, the HHA in a low herbage height and mass condition resulted in greater IGF-1 concentrations and milk yield, and induced changes in grazing and maternal behaviour that were associated with increased cows’ LW, BCS and calves’ weight at the end of the study.